From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Path: biosci!daresbury!lyra.csx.cam.ac.uk!moose.bioc.cam.ac.uk!user From: nef1002@cus.cam.ac.uk (Nick Fisher) Newsgroups: bionet.molbio.proteins Subject: Re: Proteins which run high on SDS-PAGE Date: Thu, 02 May 1996 14:19:48 +0100 Organization: Dept. of Biochemistry, University of Cambridge Lines: 22 Message-ID: References: <4m5jkf$gff@freenet-news.carleton.ca> <3187B8DF.7650@ibex.ca> NNTP-Posting-Host: moose.bioc.cam.ac.uk In article <3187B8DF.7650@ibex.ca>, Achim Recktenwald wrote: > Matt Parker wrote: > > > > Hi! We are studying a protein of 33 kDa, which consistently runs > > as about 39 kDa on SDS-PAGE (after boiling 3 mins in SDS buffer). We also > > are studying a fragment of this protein, of about 19 kDa, which runs at > > about 25 kDa. Does anybody have any clues as to why this is going on? > > > Could it be glycosylated ? On average protiens bind about 1.4g of SDS per gram > protein. Glycosylated ones bind less, that means they are less charged, they are > running as higher molecular weights. > > Achim glycoproteins tend to give very fuzzy,indistinct, microheterogeneous bands. The glycans tend to inhibit Coomassie and silver binding too. -- From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Path: biosci!ihnp4.ucsd.edu!munnari.OZ.AU!metro!metro!socs.uts.edu.au!ib_lc630.bio.uts.edu.au!smasina From: Slavica Masina Newsgroups: bionet.molbio.proteins Subject: Signal peptide sequence (corrected) Date: 3 May 1996 04:02:04 GMT Organization: University of Technology, Sydney Australia. Lines: 8 Distribution: world Message-ID: <4mc0fs$3oh@woodstock.socs.uts.EDU.AU> NNTP-Posting-Host: 138.25.241.85 X-Newsreader: Nuntius Version 1.2 X-XXMessage-ID: X-XXDate: Fri, 3 May 1996 22:08:43 GMT Does anybody know any methods, preferably recent, for the identification of protein secretory sequences and for prediciting the site of cleavage between a signal sequence and the mature exported protein. I currently have the paper of G. von Heljne (1986) Nucleic acids research, 14(11), but I would prefer computer program that performs the analysis. Can anyone help? Thanks in advance, S. Masina. From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Path: biosci!ihnp4.ucsd.edu!munnari.OZ.AU!metro!metro!socs.uts.edu.au!ib_lc630.bio.uts.edu.au!smasina From: Slavica Masina Newsgroups: bionet.molbio.proteins Subject: Signal peptide sequence Date: 3 May 1996 03:59:35 GMT Organization: University of Technology, Sydney Australia. Lines: 8 Distribution: world Message-ID: <4mc0b7$3oh@woodstock.socs.uts.EDU.AU> NNTP-Posting-Host: 138.25.241.85 X-Newsreader: Nuntius Version 1.2 X-XXMessage-ID: X-XXDate: Fri, 3 May 1996 22:06:14 GMT Does anybody know any methods, preferably recent, for the identification of protein secretory sequences and for prediciting the site of cleavage between a signal sequence and the mature exported protein. I currently have the paper of G. von Heljne (1986) Nucleic acids research, 14(11), but I would prefer computer program that performs the analysis. Can anyone help? Thanks in advance, S. Masina. From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: Looking for a DNA/Protein signal Database and Program.. Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Fri, 3 May 1996 06:39:56 GMT Lines: 14 Hello, I am trying to locate a database of signal sequences and motifs for DNA and/or Amino Acids. I would like to also locate a MAC or PC program that allows one to create a database of sequences which can then be searched for these signals. If anyone one knows where I could find such a program (we have Lasergene's Geneman, but it doesn't yet allow one to create custom databases using one's own sequences). Thanks for the help! Chuck oravaxcm@world.std.com From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nntp.coast.net!harbinger.cc.monash.edu.au!news.rmit.EDU.AU!news.unimelb.EDU.AU!munnari.OZ.AU!news.ecn.uoknor.edu!news.cis.okstate.edu!usenet From: Ron Tate Newsgroups: bionet.molbio.proteins,bionet.cellbiol Subject: Re: Product binding sites on proteins Date: Thu, 02 May 1996 15:40:03 -0500 Organization: Oklahoma State University, Biochemistry and Molecular Biology Lines: 29 Message-ID: <31891DA3.445D@bmb-fs1.biochem.okstate.edu> References: <4m5n63$73q@news.duke.edu> NNTP-Posting-Host: firefly3.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01 (Win95; I) Xref: biosci bionet.molbio.proteins:7766 bionet.cellbiol:4588 William P. Tschantz wrote: > > Hi > > I am looking for examples of proteins which have two product binding > sites (the same product) if such a protein exists. For example a protein > which has one binding site where the product is formed and then is > transferred to a second site on the enzyme. I know this sounds kind of > weird but i am trying to make sense of the data i have. The only way i > can explain it is if there is a second site present on the enzyme. I can > think of no examples except for ribosomes, which is not quite what i am > looking for. > > Any help is greatly appreciated. > > Thanks > > Bill Check out Pyruvate Dehydrogenase or any of the alpha-keto dehydrogenases. -- >>>>>>---------------------------> >Ron Tate >Lab of Franklin Leach >Dept. of Biochem. & Molecular Biology >Oklahoma State University rtate@bmb-fs1.biochem.okstate.edu (405) 744-9326 --------------------------------------------- From owner-proteins@net.bio.net Wed May 01 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!news.sprintlink.net!moonbeam.aecom.yu.edu!usenet From: auster@129.98.1.4 (auster) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Protein Elution from Nylon Date: 2 May 1996 19:08:03 GMT Organization: Albert Einstein College of Medicine Lines: 5 Message-ID: <4mb16j$i36@moonbeam.aecom.yu.edu> NNTP-Posting-Host: metatarsal.aecom.yu.edu X-Posted-From: InterNews 1.0.7@129.98.41.152 X-Authenticated: auster on POP host 129.98.1.4 Xref: biosci bionet.molbio.methds-reagnts:43975 bionet.molbio.proteins:7765 Do you know of a good protocol for eluting a protein that has been transfered by Western blot to a nylon membrane? If so, please respond to auster@aecom.yu.edu Thank you!!! From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!daresbury!hgmp.mrc.ac.uk!sanger.ac.uk!pmr From: pmr@sanger.ac.uk (Peter Rice) Newsgroups: bionet.molbio.proteins Subject: Re: Signal peptide sequence (corrected) Date: 03 May 1996 09:48:28 GMT Organization: The Sanger Centre Lines: 27 Distribution: world Message-ID: References: <4mc0fs$3oh@woodstock.socs.uts.EDU.AU> NNTP-Posting-Host: unst.sanger.ac.uk In-reply-to: Slavica Masina's message of 3 May 1996 04:02:04 GMT In article <4mc0fs$3oh@woodstock.socs.uts.EDU.AU> Slavica Masina writes: > Does anybody know any methods, preferably recent, for the identification > of protein secretory sequences and for prediciting the site of cleavage > between a signal sequence and the mature exported protein. I currently > have the paper of G. von Heljne (1986) Nucleic acids research, 14(11), > but I would prefer computer program that performs the analysis. Can > anyone help? There is a program SIGCLEAVE in EGCG (extended GCG), available from ftp.sanger.ac.uk in directory pub/pmr/egcg81 (or pub/pmr/egcg81vms for a VMS savset distribution). EGCG runs on Unix and VMS systems, and *requires* the GCG Wisconsin package. SIGCLEAVE implements Gunnar von Heijne's method, as slightly modified in his excellent book "Sequence Analysis in Molecular Biology: Treasure Trove or Trivial Pursuit". There was also an implementation of the method in PC-GENE .... -- ------------------------------------------------------------------------ Peter Rice | Informatics Division E-mail: pmr@sanger.ac.uk | The Sanger Centre Tel: (44) 1223 494967 | Hinxton Hall, Hinxton, Fax: (44) 1223 494919 | Cambs, CB10 1RQ URL: http://www.sanger.ac.uk/~pmr/ | England From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!nntp.primenet.com!news.sprintlink.net!paperboy.owt.com!news From: infomedx@oneworld.owt.com (infomedx) Newsgroups: bionet.molbio.proteins Subject: FS: Atlas of Protein Sequences and Structure 1972 Date: 3 May 1996 16:00:39 GMT Organization: One World Telecommunications Lines: 14 Message-ID: <4mdaj7$mvh@paperboy.owt.com> NNTP-Posting-Host: pm1-s9.owt.com Mime-Version: 1.0 X-Newsreader: WinVN 0.99.2 I have a hard back, ex-library copy of _Atlas of Protein Sequence and Structure 1972 Volume 5_ by Margaret O. Dayhoff in very good condition for sale. (a few stamps, but no cards, ripped out pages, etc.) Published by National Biomedical Research Foundation. The book contains 124 pages of preliminary text and 418 data pages (numbered D-1 through D-418; contains protein sequences and tables) including 20 large foldouts of sequences and structures.....$25 plus postage. --Gail infomedx@oneworld.owt.com Richland, WA USA From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!UMBI.UMD.EDU!collins From: collins@UMBI.UMD.EDU ("John H. Collins") Newsgroups: bionet.molbio.proteins Subject: Re: FS: Atlas of Protein Sequences and Structure 1972 Date: 3 May 1996 11:37:00 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <4mdaj7$mvh@paperboy.owt.com> NNTP-Posting-Host: net.bio.net I didn't realize this was a collector's item. Do you also have the three supplements? -JHC On 3 May 1996, infomedx wrote: I have a hard back, ex-library copy of _Atlas of Protein Sequence and Structure 1972 Volume 5_ by Margaret O. Dayhoff in very good condition for sale. (a few stamps, but no cards, ripped out pages, etc.) Published by National Biomedical Research Foundation. The book contains 124 pages of preliminary text and 418 data pages (numbered D-1 through D-418; contains protein sequences and tables) including 20 large foldouts of sequences and structures.....$25 plus postage. --Gail infomedx@oneworld.owt.com Richland, WA USA From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!WESLEYAN.EDU!wbeckwith From: wbeckwith@WESLEYAN.EDU (Chip) Newsgroups: bionet.molbio.proteins Subject: Protein Prep Courses? Date: 3 May 1996 08:44:47 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <3189E2D3.62F1@mail.wesleyan.edu> Reply-To: wbeckwith@wesleyan.edu NNTP-Posting-Host: net.bio.net I'm looking to better familiarize myself with some of the modern and classical methods of protein purification. I was wondering if anyone knew of any short courses in protein preparation offered sometime this summer preferably in the NE US. Any info will be greatly appreciated. Thanks William Beckwith Grad Student Wesleyan University Middletown CT wbeckwith@wesleyan.edu From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!rutgers!csn!news-1.csn.net!torn!nott!cunews!freenet-news.carleton.ca!FreeNet.Carleton.CA!at332 From: at332@FreeNet.Carleton.CA (Matt Parker) Newsgroups: bionet.molbio.proteins Subject: Re: Proteins which run high on SDS-PAGE Date: 3 May 1996 21:33:29 GMT Organization: The National Capital FreeNet Lines: 22 Sender: at332@freenet6.carleton.ca (Matt Parker) Message-ID: <4mdu39$35g@freenet-news.carleton.ca> References: <4m5jkf$gff@freenet-news.carleton.ca> <4mddrm$jir@news.tamu.edu> Reply-To: at332@FreeNet.Carleton.CA (Matt Parker) NNTP-Posting-Host: freenet6.carleton.ca Dilip Dias (v0p8630@zeus.tamu.edu) writes: > Matt > I had the same problem with a protein that I worked and working. When I > put the question in the newsgroup the answer I got was that this could > happen due to a variety of reasons. But the most important is the amount > of prolines that the protein has. Example: p53 protein of which the > calculated mol wt is about 45kd?. so be optimistic. > cheers > Dilip > > Thanks! Our proteins do have an unusually high amount of proline. Does anybody have any suggestions about why high proline content would cause the protein to run "large" on SDS-PAGE? By the way...it's not glycosylated. Thanks to all who have sent advice! Matt From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!uwm.edu!newsspool.doit.wisc.edu!news.doit.wisc.edu!martinlab From: klenchin@macc.wisc.edu (Dima Klenchin) Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: Fri, 03 May 96 21:00:38 GMT Organization: UW-Madison Lines: 13 Message-ID: <4mds5m$8kg_001@biochem.wisc.edu> References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: martinlab.biochem.wisc.edu X-Newsreader: News Xpress Version 1.0 Beta #4 In article <96124.144926JPS144@psuvm.psu.edu>, Joe Stains wrote: ->Does anyone know of any efficient way of concentrating large volumes of ->a dilute protein (several mls) for use with SDS-PAGE? I have a couple ->of ideas on some techniques, but would be interested in anything anyone ->else may have, as they all have a lot of pros and cons! I use centicones after adding SDS to the sample to 0.1% and then concentrating > 10 fold. After concentration is done, don't forget to vortex centricon REALLY well. - Dima From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!rutgers!gatech!udel!gvls1!news.cse.psu.edu!news.cac.psu.edu!psuvm!jps144 Organization: Penn State University Date: Fri, 3 May 1996 14:49:26 EDT From: Joe Stains Message-ID: <96124.144926JPS144@psuvm.psu.edu> Newsgroups: bionet.molbio.proteins Subject: Concentration of dilute proteins for SDS-PAGE Lines: 7 Does anyone know of any efficient way of concentrating large volumes of a dilute protein (several mls) for use with SDS-PAGE? I have a couple of ideas on some techniques, but would be interested in anything anyone else may have, as they all have a lot of pros and cons! Thanks in advance Joe Stains From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!rutgers!gatech!newsfeed.internetmci.com!howland.reston.ans.net!vixen.cso.uiuc.edu!newsfeed.ksu.ksu.edu!news.cis.okstate.edu!usenet From: Ron Tate Newsgroups: bionet.molbio.proteins Subject: Re: Protein Prep Courses? Date: Fri, 03 May 1996 11:51:46 -0500 Organization: Oklahoma State University, Biochemistry and Molecular Biology Lines: 22 Message-ID: <318A39A2.4660@bmb-fs1.biochem.okstate.edu> References: <3189E2D3.62F1@mail.wesleyan.edu> NNTP-Posting-Host: firefly3.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01 (Win95; I) Chip wrote: > > I'm looking to better familiarize myself with some of the modern and > classical methods of protein purification. I was wondering if anyone > knew of any short courses in protein preparation offered sometime this > summer preferably in the NE US. Any info will be greatly appreciated. > > > > > > It's not in the NE US its in Oklahoma, but our core facility at OSU is putting on just such a course June 22-26. contact Dr. Steve White (405)744-6191. -- >>>>>>---------------------------> >Ron Tate >Lab of Franklin Leach >Dept. of Biochem. & Molecular Biology >Oklahoma State University rtate@bmb-fs1.biochem.okstate.edu (405) 744-9326 --------------------------------------------- From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!ns1.faseb.org!lamarck.sura.net!newsfeed.internetmci.com!howland.reston.ans.net!torn!nott!cunews!freenet-news.carleton.ca!FreeNet.Carleton.CA!at332 From: at332@FreeNet.Carleton.CA (Matt Parker) Newsgroups: bionet.molbio.proteins Subject: Aggregation of His-tagged proteins Date: 3 May 1996 21:40:04 GMT Organization: The National Capital FreeNet We have a protein which has a six-histidine "tag" attached to the Lines: 15 Sender: at332@freenet6.carleton.ca (Matt Parker) Message-ID: <4mdufk$37u@freenet-news.carleton.ca> Reply-To: at332@FreeNet.Carleton.CA (Matt Parker) NNTP-Posting-Host: freenet6.carleton.ca N-terminal, to allow purification via a nickel-binding affinity column. We are having difficulty in concentrating this protein after purification; it tends to aggregate above 2 or 3 mg/mL. Since we are doing the concentrating at pH 7.6, I suspect that the histidines (which should be mostly uncharged at this pH) are forming a hydrophobic surface which promotes aggregation. I plan to try concentrating it at a lower pH (say 5.5 or so) to try to get some repulsion going. Has anybody had a similar problem, and can anyone offer suggestions? Thanks, Matt From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!newsfeed.internetmci.com!info.ucla.edu!library.ucla.edu!news.ucdavis.edu!usenet From: Lyle Najita Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: Fri, 03 May 1996 15:28:24 -0700 Organization: University of California, Davis Lines: 24 Message-ID: <318A8888.4B12@wheel.dcn.davis.ca.us> References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: plpstudent7.ucdavis.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01Gold (WinNT; I) Joe Stains wrote: > > Does anyone know of any efficient way of concentrating large volumes of > a dilute protein (several mls) for use with SDS-PAGE? I have a couple > of ideas on some techniques, but would be interested in anything anyone > else may have, as they all have a lot of pros and cons! > > Thanks in advance > > Joe StainsMy personal favorite if you aren't particular about recovering active protein after the gel run is to precipitate the sample with 0.1x vol. of 100% TCA solution. Mix and let sit for 20 min. to O/N and then pellet. Rinse the pellet with cold acetone to remove the residual acid. You'll know upon addition of sample buffer with BPB whether you rinsed thoroughly or not. Be sure not to let the pellet get too dry, ie - don't speed-vac the sample under heat for long periods of time, you may have problems resuspending the pellet. Lyle Najita Plant Pathology University of California - Davis From owner-proteins@net.bio.net Thu May 02 23:00:00 1996 Path: biosci!rutgers!csn!news-1.csn.net!torn!nott!bcarh189.bnr.ca!nrchh45.rich.nt.com!news.utdallas.edu!news.tamu.edu!news From: Dilip Dias Newsgroups: bionet.molbio.proteins Subject: Re: Proteins which run high on SDS-PAGE Date: 3 May 1996 16:56:22 GMT Organization: Texas A&M University Lines: 10 Message-ID: <4mddrm$jir@news.tamu.edu> References: <4m5jkf$gff@freenet-news.carleton.ca> NNTP-Posting-Host: 128.194.184.223 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:4m5jkf$gff@freenet-news.carleton.ca Matt I had the same problem with a protein that I worked and working. When I put the question in the newsgroup the answer I got was that this could happen due to a variety of reasons. But the most important is the amount of prolines that the protein has. Example: p53 protein of which the calculated mol wt is about 45kd?. so be optimistic. cheers Dilip From owner-proteins@net.bio.net Fri May 03 23:00:00 1996 Path: biosci!agate!spool.mu.edu!news.sol.net!uwm.edu!lll-winken.llnl.gov!nntp.coast.net!torn!newsflash.concordia.ca!feed.umontreal.ca!alize.ERE.UMontreal.CA!fortinfl From: fortinfl@ERE.UMontreal.CA (Fortin Flechere) Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: 4 May 96 17:14:28 GMT Organization: Universite de Montreal Lines: 16 Distribution: world Message-ID: References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: alize.ere.umontreal.ca Joe Stains writes: >Does anyone know of any efficient way of concentrating large volumes of >a dilute protein (several mls) for use with SDS-PAGE? I have a couple >of ideas on some techniques, but would be interested in anything anyone >else may have, as they all have a lot of pros and cons! >Thanks in advance >Joe Stains What I usually do to concentrate my proteins, is to precipitate them with 3 volumes of ice-cold 100% acetone. Leave on ice for 10 minutes, then spin 10 minutes at 10 000 g. Discard the supernatant. Brief wash with 80 % acetone. Spin again and let dry the pellet for 15-20 minutes. Resuspend your proteins in the volume of your choice with PBS or else. You can also use those concentrators from the company amicon or everything like that, but it can take several hours to obtain what you whant in volume. Ho pe it will help you. Flechere Fortin ZZ From owner-proteins@net.bio.net Fri May 03 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!nntp.coast.net!howland.reston.ans.net!swrinde!newsfeed.internetmci.com!in1.uu.net!news.puc.cl!news From: bvgarret@genes.bio.puc.cl (Virginia) Newsgroups: bionet.molbio.proteins Subject: How do can I find protein domains? Date: 4 May 1996 16:32:03 GMT Organization: PUC-Chile Lines: 9 Message-ID: <4mg0q3$juk@huequi.puc.cl> NNTP-Posting-Host: xjordana2.bio.puc.cl Mime-Version: 1.0 Content-Type: Text/Plain; charset=US-ASCII X-Newsreader: WinVN 0.99.7 Help!!! I am train to see if the protein I am studiying ( a transcription factor) has a Casein Kinase fosforilation domain. Some body knows how can I do so? Hint: I have the sequence. Thanks Virginia From owner-proteins@net.bio.net Fri May 03 23:00:00 1996 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!newsfeed.ksu.ksu.edu!usenet From: scotbean@ksu.ksu.edu (Scott Bean) Newsgroups: bionet.molbio.proteins Subject: Phosphorylated proteins and CE Date: 4 May 1996 16:25:57 GMT Organization: kansas state university Lines: 14 Message-ID: <4mg0el$lol@newserv.ksu.ksu.edu> NNTP-Posting-Host: pc31gqu.usgmrl.ksu.edu Mime-Version: 1.0 Content-Type: Text/Plain; charset=US-ASCII X-Newsreader: WinVN 0.99.7 Has anyone done any analysis of phosphorylated proteins using capillary electrophoresis or know of any references to papers that have separated phosphorylated proteins using CE? I'm looking for a method to look at the microhetergeneity (sp?) that might be caused by phosphorylation. Can you separate "phosphoforms"? Also, has any done or know of the same thing except done with reversed-phase HLPLC? Thanks. Scott Bean From owner-proteins@net.bio.net Fri May 03 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!vixen.cso.uiuc.edu!uwm.edu!newsspool.doit.wisc.edu!news.doit.wisc.edu!martinlab From: klenchin@macc.wisc.edu (Dima Klenchin) Newsgroups: bionet.molbio.proteins Subject: Re: Phosphorylated proteins and CE Date: Sat, 04 May 96 17:15:44 GMT Organization: UW-Madison Lines: 18 Message-ID: <4mg3c0$8kg_001@biochem.wisc.edu> References: <4mg0el$lol@newserv.ksu.ksu.edu> NNTP-Posting-Host: martinlab.biochem.wisc.edu X-Newsreader: News Xpress Version 1.0 Beta #4 In article <4mg0el$lol@newserv.ksu.ksu.edu>, scotbean@ksu.ksu.edu (Scott Bean) wrote: ->Has anyone done any analysis of phosphorylated proteins using capillary ->electrophoresis or know of any references to papers that have separated ->phosphorylated proteins using CE? I'm looking for a method to look at the ->microhetergeneity (sp?) that might be caused by phosphorylation. Can you ->separate "phosphoforms"? -> ->Also, has any done or know of the same thing except done with reversed-phase ->HLPLC? I have no knowledge/experience with CEF, but as far as "phospho-isoforms" are concerned, my method of choice would be chromatofocusing. Guys in our lab were able to separate 0,1,2,3...-phosphorylated forms of rhodopsin on Pharmacia's Mono-P column. (It's expensive, but you don't have to buy FPLC to run it - regular HPLC setup works just fine). - Dima From owner-proteins@net.bio.net Fri May 03 23:00:00 1996 Path: biosci!ns1.faseb.org!lamarck.sura.net!newsfeed.internetmci.com!sgigate.sgi.com!news.cs.indiana.edu!news.nstn.ca!newsflash.concordia.ca!news.mcgill.ca!news From: dasuser@PO-Box.McGill.CA Newsgroups: bionet.molbio.proteins Subject: Post Doc on Neurofibromatosis type 2 protein Date: 4 May 1996 21:58:32 GMT Organization: McGill University Computing Centre Lines: 26 Message-ID: <4mgju8$ljm@sifon.cc.mcgill.ca> NNTP-Posting-Host: d-10.das.mcgill.ca X-Newsreader: SPRY News 3.03 (SPRY, Inc.) **Please DO NOT forward your application via e-mail.*** ======== A postdoctoral position is available in the laboratory of Dr. Guy A. Rouleau at the Montreal General Hospital. The postdoctoral fellow will work on the cell biology of the Neurofibromatosis type 2 protein. Applicants should have experience in cell culture and transfection, cell and molecular biology techniques, and protein analysis. The postdoctoral fellow should be creative, independent and strongly motivated. The postdocotral position is available as soon as possible. Please send your curriculum vitae and statement of research interest to Guy A. Rouleau, M.D., Ph.D. Center for Research in Neuroscience, Montreal General Hospital, Montreal, Quebec, H3G 1A4. **Please DO NOT forward your application via e-mail.*** ======== From owner-proteins@net.bio.net Sat May 04 23:00:00 1996 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!newsfeed.internetmci.com!xmission!news.cc.utah.edu!news.cs.utah.edu!swen.emba.uvm.edu!hbeernin From: hbeernin@med.uvm.edu (Hans Beernink) Newsgroups: bionet.molbio.proteins Subject: Re: Protein folding Date: 2 May 1996 19:45:52 GMT Organization: EMBA Computer Facility, The University of Vermont Lines: 41 Message-ID: <4mb3dg$3s7@swen.emba.uvm.edu> References: NNTP-Posting-Host: protein.med.uvm.edu X-Newsreader: TIN [version 1.2 PL1] David Micklem (drm21@mole.bio.cam.ac.uk) wrote: : What I have is the NMR structure of a particular protein domain. I also : have the sequence of several (many) similar domains which show a high : degree of sequence homology to the one whose structure has been : determined. What I want to do is to see how well these homologues 'fit' : the determined structure. If this could be used as the basis for a search : for a 'local' most-likely stable structure, then so much the better. Of : course, if the sequence provided _couldn't_ stably form the template : structure, or anything close, that would be interesting too. And if it : could interpret the results and write up a couple of papers for me : too.... David- it sounds like you need to do some molecular dynamics. There are some (relatively) user friendly packages (e.g. insight with the discover module, xplor etc.) that will allow you to "thread" your AA sequence into a structure coordinate file. After minimizing your structure, you can compare the results with the original "anchor" structure and make some predictions. Best regards, Hans -- _____________________________________________________________________________ "The worst monotonous drone coming from a lectern or the most eye-splitting textbook written in turgid English is nothing in comparison to the psychological Sahara that starts right in your bedroom and spurns the horizon." -Joseph Brodsky, from "In praise of Boredom" delivered as a commencement address at Dartmouth College. _____________________________________________________________________________ Hans T.H. Beernink, Department of Biochemistry, University of Vermont hbeernin@protein.med.uvm.edu FAX (802)862-8229 hbeernin@zoo.uvm.edu Tel.(802)656-8244 URL http://moose.uvm.edu/~hbeernin/ URL http://salus.med.uvm.edu/biochem/ From owner-proteins@net.bio.net Sat May 04 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!newsfeed.internetmci.com!in2.uu.net!brighton.openmarket.com!decwrl!newsspool.doit.wisc.edu!news.doit.wisc.edu!martinlab From: klenchin@macc.wisc.edu (Dima Klenchin) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: Sun, 05 May 96 22:55:18 GMT Organization: UW-Madison Lines: 23 Message-ID: <4mjbkm$6is_001@biochem.wisc.edu> References: NNTP-Posting-Host: martinlab.biochem.wisc.edu X-Newsreader: News Xpress Version 1.0 Beta #4 Xref: biosci bionet.general:21490 bionet.biology.cardiovascular:937 bionet.cellbiol:4607 bionet.glycosci:658 bionet.biophysics:1936 bionet.molbio.ageing:2686 bionet.molbio.proteins:7788 bionet.neuroscience:13949 sci.med:122383 sci.research.careers:10114 talk.politics.medicine:51169 In article , Bert Gold wrote: Newsgroups: >bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bi >onet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience >,sci.med,sci.research.careers,talk.politics,talk.politics.medicine >My hope in writing this is that our children will not suffer >because of the lack of wisdom of their leaders in >making decision about what to and what not to study. Judging by the number of newsgroups crossposted, Bert Gold fully qualifies as spammer... :-)) Just kidding Seriously: Dr. Gold has obviously very strong opinion on the subject. The Q: is it really confirmed by research, or the current state of affair is simply that kilogram amounts of ascorbic acid don't hurt (apparently)? - Dima From owner-proteins@net.bio.net Sat May 04 23:00:00 1996 Path: biosci!agate!cgl!itssrv1.ucsf.edu!itsa.ucsf.edu!bgold From: Bert Gold Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: Sun, 5 May 1996 08:36:12 -0700 Organization: UCSF, ITS Lines: 132 Message-ID: NNTP-Posting-Host: itsa.ucsf.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII To: Dixie Lawrence cc: John Burris Xref: biosci bionet.general:21489 bionet.biology.cardiovascular:936 bionet.cellbiol:4605 bionet.glycosci:657 bionet.biophysics:1935 bionet.molbio.ageing:2685 bionet.molbio.proteins:7787 bionet.neuroscience:13947 sci.med:122374 sci.research.careers:10113 talk.politics.medicine:51166 April 22, 1996 Walking back from the bank to the hospital today I was prompted to buy several large juicy navel oranges. I've been eyeing oranges for many weeks now, but my wife and I have a long standing disagreement about who picks the best ones. I thought the oranges she chose at a fruit stand, just outside Half Moon Bay yesterday, were puny; she again suggested I always pick the oversized, dried up ones. Today, though, my oranges were winners: Sweet, juicy and large; just the way I like them. In the news the past several days, as you've probably heard, Vitamin C requirements (RDAs, Required Daily Allowance, set by the National Research Council, the nation's highest scientific authority) for adult males have been revalued upward. Not modestly either: But three-fold, and still the upper limit is poorly defined. It seems an intramural researcher at NIH finally got the funding go ahead to do some of the experiments which Linus Pauling and Albert Szent-Gyorgyi always longed to do before the end of their lives, but were denied funding for. It is notable that in this latest work (1), seven men in their twenties were fed increasing doses of Vitamin C after being starved for it in their diet. Although they never showed signs of scurvy, each volunteer did report feeling uncomfortable when deprived of Vitamin C. The NIH authors note that because the subjects of their study were young men in their 20's, the results are limited to this subject group. The vivid image of Szent-Gyorgyi standing in front of Whitman Auditorium in Woods Hole holding a bottle of glyoxyl in one hand and Vitamin C in the other is permanently etched in my memory. I'll not easily forget the hypothesis of 'radical scavenging' which Szent-Gyorgyi put to us that day: He said that he and Pauling had been talking about the central importance of OXYGEN in human metabolism and explained that he had been concerned for a good deal of time about reactions generating singlet oxygen. These, produced in abundance during mitochondrial "charge transfer" in liver parenchyma, and other aerobically active tissues, must absorb electrons pairwise in order to avoid causing damage. In 1931, Pauling discovered the superoxide radical, which, though produced in minute quantities as an unwanted byproduct of oxidative phosphorylation, has enormous destructive capacity if not defused (2). During lectures given at UC Berkeley in the 80s, Pauling paid tribute to the remarkable specificity of the enzymatic system that neutralizes it: Superoxide dismutase; at the same time, however, he noted that this enzyme does not defuse the lesser, but still significant intracellular destructive capacity of singlet oxygen or the hydroxide radical (and unkown to him at the time, the NO radical). Superoxide dismutase transforms superoxide radicals to peroxide which can then be eliminated as water after the action of peroxidase and catalase. But what is the body to do with all the unpaired electrons such a process would generate? Szent-Gyorgyi told that oxygen itself could absorb unpaired electrons if only it were conjugated to carbon in the form of a double carbonyl: the simplest such compound being glyoxyl, commonly found in human liver, for this compound had sufficient electron delocalization to provide something of an electron sink. And, the supply of glyoxyl is renewable through the action of the glutathione-S-tranferase system, present in liver mitochondria. Further, Szent-Gyorgyi told us excitedly, two englishmen and a german, H.D. Dakin, H.W. Dudley and C. Neuberg had, in 1913 discovered that methylated-glyoxyl could be transformed to a potent energy source itself, lactic acid, through the action of glyoxylase, which they discovered in that year. So, in one fell swoop, Szent-Gyorgyi had connected for us the relationship between oxygen radical and one carbon metabolism. I promise I will discuss this relationship in the next few weeks, when I write another essay, tentatively titled, Folic Acid and I. In comments honoring Szent-Gyorgyi on the occasion of his 82nd birthday, Linus Pauling recalled Vitamin C's discovery by Szent-Gyorgyi in 1928. Pauling noted that Szent-Gyorgyi had written in 1939 (4) that although organisms were generally well adapted to their surroundings, that the destruction of the natural environment endangered that adapatation. "I have a strong faith in the perfection of the human body", Szent-Gyorgyi wrote in 1939, "and I think that vitamins are an important factor in its coordination with its surroundings. Vitamins, if properly understood and applied, will help us to reduce human suffering to an extent which the most fantastic mind would fail to imagine." (3) On the day that I met him in Woods Hole, Szent-Gyorgyi told us he and Pauling (three Nobel prizes) between them suggested that significant increases in Vitamin C intake were almost certainly required by an adult body, perhaps especially at times when oxygen production was increased. Szent-Gyorgyi had insisted in a book he had written two years before, that Vitamin C, especially when complexed with manganese in the presence of oxygen, could synthesize a free radical form, as well and as easily as it could form a dehydroascorbate, oxidized form in the presence of copper or iron and in alliance with an enzyme he'd discovered in plants in 1931 and named 'ascorbic acid oxidase.' Szent-Gyorgyi insisted that it was in some sense the equilibrium between the various oxidized and reduced forms of ascorbate which provided its multifarious activities. And yet, this week, fully 18 years after the memorable talk by Szent-Gyorgyi which I just described, the Vitamin C RDA for men has been adjusted three-fold upward. It makes me wonder at the brazen inefficiency of the research 'establishment'... The knowledge that both Linus and Albe died without ever attaining a modicum of funding for their final nutrition project confirms some of my worst fears about our ways of deciding what research is worthy in this country. My hope in writing this is that our children will not suffer because of the lack of wisdom of their leaders in making decision about what to and what not to study. Bert Gold San Francisco References (1) Levine, M.; Conry-Cantilena, C.; Wang, Y.; Welch, R.W.; Washko, P.W.; Dhariwal, K.R.; Park, J.B.; Lazarev, A.; Graumlich, J.G., King, J.; Cantilena, L.R. (1996) Vitamin C pharmacokinetics in healthy volunteers: Evidence for a recommended dietary allowance. Proc. Natl. Acad. Sci. USA; 93, 3704-3709. (2) Marinacci, B. (1995) Linus Pauling in His Own Words, New York, Simon & Shuster. (3) Kaminer, B., ed. (1977) Search and Discovery, A tribute to Albert Szent-Gyorgyi, New York, Academic Press. (4) Szent-Gyorgyi, A. (1939) On Oxidation, Fermentation, Vitamins, Health and Disease. Baltimore, Williams and Wilkins. From owner-proteins@net.bio.net Sat May 04 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!nntp.coast.net!howland.reston.ans.net!newsfeed.internetmci.com!in2.uu.net!brighton.openmarket.com!decwrl!newsspool.doit.wisc.edu!news.doit.wisc.edu!martinlab From: klenchin@macc.wisc.edu (Dima Klenchin) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Q: Properties of Phospho-cellulose (P11, Whatman) Date: Sun, 05 May 96 23:13:18 GMT Organization: UW-Madison Lines: 20 Message-ID: <4mjcme$6j8_001@biochem.wisc.edu> NNTP-Posting-Host: martinlab.biochem.wisc.edu X-Newsreader: News Xpress Version 1.0 Beta #4 Xref: biosci bionet.molbio.methds-reagnts:44083 bionet.molbio.proteins:7789 Dear all, I'm using P11 as a first step of protein purification. In a little booklet that comes with it, the company puts some pretty scary notes about it's use. Something like "precycle, use right away or store no more than a week, and only in no less than 0.5 M phosphate". Sort of justification for it is alledged instability of ester linkage to the cellulose. My Q is: is this really truth? Does phosphocellulose really go bad over prolonged storage? I mean, it't kind of pain in the neck to precycle it. Life would so much easier if can store it in ready-to-use form for at least month. Can I simply add phosphate, azide and count on the same results with "old" stuff as with freshly prepared matrix? Folks experienced with phosphocellulose, please, send your comments. Thanks in advance, - Dima From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!ihnp4.ucsd.edu!scripps.edu!NewsWatcher!user From: anonymous@scripps.edu (Anonymous) Newsgroups: bionet.molbio.proteins Subject: 2-Dgels Date: Mon, 06 May 1996 10:01:51 -0700 Organization: The Scripps Research Institute Lines: 8 Message-ID: NNTP-Posting-Host: mahler.scripps.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit I am in the process of selecting a system to run 2-D gels. Any thoughts as to what is the best system to use and what I should be aware of as I have never done this before. Any good, current references on the subject? Can respond via email to: ranheim@scripps.edu Thanks From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!surfnet.nl!swsbe6.switch.ch!scsing.switch.ch!news.belwue.de!news.dfn.de!uni-muenster.de!news From: schille@uni-muenster.de Newsgroups: bionet.molbio.proteins Subject: S: tryptase Date: 6 May 1996 14:39:43 GMT Organization: Westfaelische Wilhelms-Universitaet Muenster, Germany Lines: 5 Message-ID: <4ml2vf$tua@majestix.uni-muenster.de> NNTP-Posting-Host: fplc.uni-muenster.de X-Newsreader: AIR News 3.X (SPRY, Inc.) Does anyone know from whom we can obtain the enzyme tryptase ( human or bovine)? Please mail us, if you know. Thanks for your help. e-mail: kusters@uni-muenster.de From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!galaxy.ucr.edu!library.ucla.edu!agate!cgl!itssrv1.ucsf.edu!itsa.ucsf.edu!bgold From: bgold@itsa.ucsf.edu (Bert Gold) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Followup-To: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Date: 6 May 1996 12:37:24 GMT Organization: UCSF, ITS Lines: 22 Message-ID: <4mkrq4$pih@itssrv1.ucsf.edu> References: <4mjbkm$6is_001@biochem.wisc.edu> NNTP-Posting-Host: itsa.ucsf.edu X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.general:21501 bionet.biology.cardiovascular:939 bionet.cellbiol:4610 bionet.glycosci:661 bionet.biophysics:1937 bionet.molbio.ageing:2688 bionet.molbio.proteins:7793 bionet.neuroscience:13956 sci.med:122444 sci.research.careers:10122 talk.politics.medicine:51190 Dima, My reading of the aforemented PNAS paper by Levine et al., (1996) disagrees with the very high dose vitamin C protocols which were advocated by Pauling and Szent-Gyorgyi, without providing any firm evidence on the issue, and without mentioning these great investigators by name. Conclusions in the PNAS paper are limited to a new RDA for ascorbate for men in their 20s. Bert Gold San Francisco Dima Klenchin (klenchin@macc.wisc.edu) wrote: : Q: is it really confirmed by research, or the current state of affair : is simply that kilogram amounts of ascorbic acid don't hurt (apparently)? : - Dima From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!usc!hookup!uwm.edu!lll-winken.llnl.gov!enews.sgi.com!news.uoregon.edu!arclight.uoregon.edu!dispatch.news.demon.net!demon!netcom.net.uk!ix.netcom.com!news From: mhaha@ix.netcom.com (Glenn Diamond) Newsgroups: bionet.neuroscience,bionet.molbio.methds-reagnts,bionet.molbio.proteins,sci.chem,sci.med.pharmacy,sci.bio.biotechnology Subject: New Company Formed - Looking for People Date: 6 May 1996 10:39:23 GMT Organization: Netcom Lines: 25 Message-ID: <4mkksr$rp8@dfw-ixnews4.ix.netcom.com> NNTP-Posting-Host: min-mn2-17.ix.netcom.com X-NETCOM-Date: Mon May 06 5:39:23 AM CDT 1996 Xref: biosci bionet.neuroscience:13954 bionet.molbio.methds-reagnts:44093 bionet.molbio.proteins:7792 sci.chem:55520 sci.med.pharmacy:26745 Colleagues, This is to announce the birth yesterday of a new company focused on Peptide Neuropharmaceuticals with an Informatics-focused Drug Discovery Process. NDI is looking for people with the following backgrounds to participate as employees, consultants, or high or low level advisers: Neurobiology, neurochemistry, peptide design theory, peptide synthetic chemistry, neurochemical immunoassay, pharmaceutical animal study design, and peptide bioavailability engineering, and neuropeptide sciences. If you are interested, please provide your thoughts and your brief bio with brief biblio to mhaha@ix.netcom.com and your message will be reviewed and answered. Thank you. Glenn Diamond President NDI From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!nntp.coast.net!oleane!in2p3.fr!swidir.switch.ch!swsbe6.switch.ch!scsing.switch.ch!rzunews.unizh.ch!NewsWatcher!user From: maga@vetbio.unizh.ch (Giovanni Maga) Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: Mon, 06 May 1996 11:35:12 -0500 Organization: University of Zurich Irchel- Biochemistry Lines: 24 Message-ID: References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: 130.60.120.11 In article <96124.144926JPS144@psuvm.psu.edu>, Joe Stains wrote: > Does anyone know of any efficient way of concentrating large volumes of > a dilute protein (several mls) for use with SDS-PAGE? I have a couple > of ideas on some techniques, but would be interested in anything anyone > else may have, as they all have a lot of pros and cons! > > Thanks in advance > > Joe Stains One possible way: dialyze your sample against a buffer with a concentration of all the components which is, let's say, 10-fold less the one you have in your starting sample. Then use a speed-vac to concentrate the sample up to the starting concentration of buffer components (in this case 10-fold). The dialysis step is in order to avoid to concentrate too much whatever you have in your buffer, since it could then disturb the run on the SDS-PAGE. If the kind of buffer does not bother you, just dialyze against ammonium acetate (at the right pH for your protein) and then speed-vac it down to the volume you like (in this case you can reduce the volume to nothing, having only your protein in the lyophilized material at the end). It's just an idea...I did it and it worked. Hope the same with you. From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!PSUVM.PSU.EDU!JPS144 From: JPS144@PSUVM.PSU.EDU ("Joe Stains") Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: 6 May 1996 08:11:59 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: <199605061511.IAA17409@net.bio.net> NNTP-Posting-Host: net.bio.net Could you give me some specifics on the PEG 8000 Protein ppt? Thanks. Joe Stains From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!NMSU.EDU!hroychow From: hroychow@NMSU.EDU (HIRANYA ROYCHOWDHURY) Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: 6 May 1996 08:00:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 44 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: net.bio.net On Fri, 3 May 1996, Joe Stains wrote: > Does anyone know of any efficient way of concentrating large volumes of > a dilute protein (several mls) for use with SDS-PAGE? I have a couple > of ideas on some techniques, but would be interested in anything anyone > else may have, as they all have a lot of pros and cons! > > Thanks in advance > > Joe Stains > Tha following are a few of the may ways, without the use of commercially avilable concentrators: 1. Precipitation with 90% ammonium sulfate- brings down almost 95% of the proteins. Dialysis against a low salt buffer required. 2. Precipitation with acetone. The acetone conc. to be determined empirically. 3. Pptn with TCA. A part of the ppt. will not resolubilize, most proteins would dissolve back into Laemmli sample buffer and the resulting SDS-PAGE pattern would be fairly representative. 4. EtOH pptn. works the same way as the TCA pptn., and is less harsh. But, a good percentage of the proteins may not ppt. in alcohol. 5. PEG 8000. This is my favorite. But, the salt concentration also increases so one may need to carry out drop dialysis before preparing sample for electrophoresis. >>>>>>>>>>>>>>>>>>>>>>>>>>> Hiranya S. Roychowdhury Plant Genetic Engineering Lab. Box 3GL, NM State Univ. Las Cruces, NM 88003 Phone: (505) 646-5785 hroychow@nmsu.edu <<<<<<<<<<<<<<<<<<<<<<<<<<< From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!howland.reston.ans.net!newsfeed.internetmci.com!news.dacom.co.kr!nntp.coast.net!torn!resunix.ri.sickkids.on.ca!news From: Randall Willis Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: Q: Properties of Phospho-cellulose (P11, Whatman) Date: 6 May 1996 12:52:42 GMT Organization: The Hospital for Sick Children Lines: 30 Message-ID: <4mksmq$9l2@resunix.ri.sickkids.on.ca> References: <4mjcme$6j8_001@biochem.wisc.edu> NNTP-Posting-Host: resunix.ri.sickkids.on.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) To: klenchin@macc.wisc.edu X-URL: news:4mjcme$6j8_001@biochem.wisc.edu Xref: biosci bionet.molbio.methds-reagnts:44103 bionet.molbio.proteins:7794 Dima, Unfortunately, what Whatman says is true. I have worked with P11 off and on for over 8 years and have found it to be the most intractable resin on the market. With no shelf life and very labile active groups, the reproduceability of the resin from preparation to preparation is aggravating. My recommendation would be to find another resin. If you need a cation exchanger, try CM cellulose (weak) or sulphopropyl (SP, strong) instead. I believe that Whatman sells both and I know that Pharmacia sells the SP in a variety of resin bead types. If the phosphate is key to your binding, may I recommend that you generate your own resin. In our case, we have affixed phosphotyrosine to CNBr-activate sepharose 4B (Pharmacia) and use this to purify P-Tyr binding proteins. It's a drag and I would love it if someone could produce a good phosphorylated resin but nothing is out there of which I am aware. Good luck...Randall C Willis Publisher Biochemistry Research Aliquotes Press Hosp for Sick Children "ALIQUOTES:A Journal of Molecular 555 University Ave. and Biochemical Humour" Toronto, ON 58 Balfour Ave. M5G 1X8 CANADA Toronto, ON M4C 1T6 CANADA willis@gandalf.psf. sickkids.on.ca rogerb@microsoft.com From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!wizard.pn.com!brighton.openmarket.com!decwrl!enews.sgi.com!news.uoregon.edu!neonlights.uoregon.edu!usenet From: matt thomas Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: Q: Properties of Phospho-cellulose (P11, Whatman) Date: Mon, 06 May 1996 18:48:37 +0000 Organization: University of Oregon Lines: 26 Message-ID: <318E4985.7730@molbio.uoregon.edu> References: <4mjcme$6j8_001@biochem.wisc.edu> NNTP-Posting-Host: longhawl.uoregon.edu Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Mailer: Mozilla 2.01 (Macintosh; I; PPC) Xref: biosci bionet.molbio.methds-reagnts:44142 bionet.molbio.proteins:7800 I have used P11 for a couple of different purifications (rap74, coli RNA polymerase, yeast pol I, calf thymus pol II) and I have been able to store it up to 6 months with no change in eluttion profile or activity of the prep. I found if you store it in 200mM Tris pH 7.9 at 4°C (thats right 200 mM, it is not a typo), 1mM EDTA, and NaN3 it seemes to do ok. Leave out the glycerol so buges don't have a carbon source. You have to double check the pH before you pour the column though. Stick your pH prop right into the slury to do this. A few time I have used p11 that was older then 6 months stored under these same conditions and everythign seemed fine, but I only did this once or twice so I don't want to bet the farm on it. Is long as you check the pH and don't overload the column I have had good luck with this resgin. It does need to be babied and probably is not the best thing out there, but if it works why fix it? :) Matt Thomas Thomas@molbio.uoregon.edu Institue of molecular biology University of Oregon > > From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!news.mathworks.com!newsfeed.internetmci.com!globe.indirect.com!usenet From: "John E. Kuslich" Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: Mon, 06 May 1996 18:37:27 -0600 Organization: CRAK Software Lines: 56 Message-ID: <318E9B47.285B@indirect.com> References: NNTP-Posting-Host: s178.phxslip4.indirect.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0 (Win95; I) CC: news Xref: biosci bionet.general:21511 bionet.biology.cardiovascular:943 bionet.cellbiol:4616 bionet.glycosci:663 bionet.biophysics:1941 bionet.molbio.ageing:2690 bionet.molbio.proteins:7801 bionet.neuroscience:13968 sci.med:122540 sci.research.careers:10137 talk.politics.medicine:51213 Bert Gold wrote: > <<<>> > And yet, this week, fully 18 years after the memorable talk by > Szent-Gyorgyi which I just described, the Vitamin C RDA for men > has been adjusted three-fold upward. > > It makes me wonder at the brazen inefficiency of the research > 'establishment'... > > The knowledge that both Linus and Albe died > without ever attaining a modicum of funding for their final > nutrition project confirms some of my worst fears about > our ways of deciding what research is worthy in this > country. > > My hope in writing this is that our children will not suffer > because of the lack of wisdom of their leaders in > making decision about what to and what not to study. > > Bert Gold > San Francisco > > References <<> Just look at research on "Cold Fusion", "Global Warming", and "The Ozone Hole" for further examples of the poor quality of some recent so-called scientific research. Chaos theory eliminated any hope of long term weather prediction, and yet we still have "researchers" at major universities doing computer simulation of weather. Modern science has gone "politically correct"; look at the reaction to that book entitled "The Bell Curve". It was widely renounced by many so-called scientists who never took the time to read it!!! Oh well... John E. Kuslich -- John E. Kuslich WPcrak for Wordperfect johnk@indirect.com WDcrak for MS Word Password Recovery EXcrak for MS Excel 602 863 9274 voice QPcrak for Quattro Pro 602 548 1993 fax LOcrak for Lotus 123 Password Removal QWcrak for Quicken Our WEB Site http://www.indirect.com/www/johnk/ >>>>>>>>You Hack'em, We CRAK'em<<<<<<<<<<<<< From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Path: biosci!ns1.faseb.org!lamarck.sura.net!newsfeed.internetmci.com!uwm.edu!vixen.cso.uiuc.edu!newsfeed.ksu.ksu.edu!news.cis.okstate.edu!usenet From: Ron Tate Newsgroups: bionet.molbio.proteins Subject: Re: Concentration of dilute proteins for SDS-PAGE Date: Mon, 06 May 1996 11:58:43 -0500 Organization: Oklahoma State University, Biochemistry and Molecular Biology Lines: 23 Message-ID: <318E2FC3.1E7F@bmb-fs1.biochem.okstate.edu> References: <96124.144926JPS144@psuvm.psu.edu> NNTP-Posting-Host: firefly3.biochem.okstate.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01 (Win95; I) Joe Stains wrote: > > Does anyone know of any efficient way of concentrating large volumes of > a dilute protein (several mls) for use with SDS-PAGE? I have a couple > of ideas on some techniques, but would be interested in anything anyone > else may have, as they all have a lot of pros and cons! > > Thanks in advance > > Joe Stains Joe, I used to use the TCA method but I've switched to a phenol/ether extraction method that I got from -Analytical Biochemistry- vol.226, 1995, pp.382-383. It works really well. -- >>>>>>---------------------------> >Ron Tate >Lab of Franklin Leach >Dept. of Biochem. & Molecular Biology >Oklahoma State University rtate@bmb-fs1.biochem.okstate.edu (405) 744-9326 --------------------------------------------- From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!nntp.primenet.com!uunet!in1.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: Where can I find a database of peptidases? Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Tue, 7 May 1996 04:25:40 GMT Lines: 7 I am looking for a database of known peptidases and their recognition sites. If anyone knows where I can find this I'd be most appreciative.. Chuck oravaxcm@world.std.com From owner-proteins@net.bio.net Sun May 05 23:00:00 1996 Newsgroups: bionet.molbio.proteins Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!nntp.primenet.com!uunet!in1.uu.net!world!oravaxcm From: oravaxcm@world.std.com (Charles A Miller) Subject: Any info on Ariadne files? Message-ID: Organization: The World Public Access UNIX, Brookline, MA X-Newsreader: TIN [version 1.2 PL2] Date: Tue, 7 May 1996 04:27:49 GMT Lines: 11 Does anyone know of a resource for creating Ariadne files, or an archive of known Ariadne files. We have Lasergene and would like to be able to use these files to further characterize our sequence. Thanks! Chuck oravaxcm@world.std.com From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!IRIS.LSC.PKU.EDU.CN!lisl From: lisl@IRIS.LSC.PKU.EDU.CN (Li Songlin) Newsgroups: bionet.molbio.proteins Subject: Ebola Virus Date: 7 May 1996 21:56:51 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <9605081952.AA26024@IRIS.lsc.pku.edu.cn> NNTP-Posting-Host: net.bio.net In Message-Id: <4mnq1b$ppf@newsie.uscyber.com>, Kelly wrote, >I'm doing a Term paper on the Ebola Virus. I'd Appreciate any possible I believe that Ebola Virus is a subject for three dimmentional reconstruction. I do not remmenber who are working on it. Prof. Guangying Lu might give you some help, I believe. Songlin Li PS, Sorry. The email address of Prof. Guangying Lu is lgy@iris.lsc.pku.edu.cn From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!brighton.openmarket.com!decwrl!purdue!mozo.cc.purdue.edu!not-for-mail From: hrubin@b.stat.purdue.edu (Herman Rubin) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: 7 May 1996 20:29:21 -0500 Organization: Purdue University Statistics Department Lines: 28 Message-ID: <4motdh$3mfi@b.stat.purdue.edu> References: <318E9B47.285B@indirect.com> NNTP-Posting-Host: b.stat.purdue.edu Xref: biosci bionet.general:21530 bionet.biology.cardiovascular:949 bionet.cellbiol:4629 bionet.glycosci:668 bionet.biophysics:1947 bionet.molbio.ageing:2696 bionet.molbio.proteins:7823 bionet.neuroscience:13991 sci.med:122684 sci.research.careers:10154 talk.politics.medicine:51251 In article <318E9B47.285B@indirect.com>, John E. Kuslich wrote: >Bert Gold wrote: >> <<<>> >> And yet, this week, fully 18 years after the memorable talk by >> Szent-Gyorgyi which I just described, the Vitamin C RDA for men >> has been adjusted three-fold upward. >> It makes me wonder at the brazen inefficiency of the research >> 'establishment'... ................. >Chaos theory eliminated any hope of long term weather prediction, and >yet we still have "researchers" at major universities doing computer >simulation of weather. That we cannot predict the precise time at which rain will start, or things to that effect, does not mean that we cannot learn by judicious use of simulation. Choas theory tells us that exactly what will happen is very sensitive to initial conditions, but that we cannot tell exactly where that tornado will strike is no reason not to give a tornado warning. ................... -- Herman Rubin, Dept. of Statistics, Purdue Univ., West Lafayette IN47907-1399 hrubin@stat.purdue.edu Phone: (317)494-6054 FAX: (317)494-0558 From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!galaxy.ucr.edu!library.ucla.edu!agate!cgl!itssrv1.ucsf.edu!itsa.ucsf.edu!bgold From: bgold@itsa.ucsf.edu (Bert Gold) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Followup-To: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Date: 8 May 1996 01:15:55 GMT Organization: UCSF, ITS Lines: 91 Message-ID: <4moskb$14rs@itssrv1.ucsf.edu> References: <4mjbkm$6is_001@biochem.wisc.edu> NNTP-Posting-Host: itsa.ucsf.edu X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.general:21531 bionet.biology.cardiovascular:950 bionet.cellbiol:4630 bionet.glycosci:669 bionet.biophysics:1948 bionet.molbio.ageing:2697 bionet.molbio.proteins:7824 bionet.neuroscience:13993 sci.med:122692 sci.research.careers:10155 talk.politics.medicine:51252 Mr. Kuslich's suggestion that science can somehow be cleansed of human values and thus be made more pure is a misunderstanding of what science is and does. Science is a human endeavor. As such it manifests all of the frailties and foibles of any deeply human undertaking. It cannot be politically 'cleansed' of value: Further, why would you want to? Science, as practiced, should be ethical. It should aspire to revere its creators, not dimish them. Bronowski and Pauling wrote eloquently on these points. Pauling always suggested that the goal of science was clearly to diminish human suffering. That is why he was awarded a Nobel Prize in Peace; for helping to orchestrate the Test Ban Treaty; it is also notable that he was blacklisted, and almost indicted for treason for that very same cause. Science is political. But it should not be guided solely by a political agenda: That is why it has (for the last couple of hundred years) been done mostly at Universities, where tenure (an archaic, but important system for guarding unpopular ideas) has been guaranteed. Tenure is not perfect, witness the case of Paul Simmelweiss, who died a pauper in the streets of Vienna for having the audacity to suggest that surgeons should wash themselves between autopsy and entering the operating theatre. No, tenure is not perfect, but it is better than nothing.... We are now, in the process, in this country, of doing away with all that! Humanistic science, tenure, Universities (as sanctuaries for learning) are all being destroyed. Won't you who are criticizing the blind alleys into which science inevitably goes admit your own imperfection: So that you can help to save a tradition of inquiry which, although imperfect, used to be credible, ethical and respectable. That was my goal in writing this essay: To acknowledge that we have lost something if we do not clearly respect the humility of our own enterprise of discovery. And perhaps the most important sense of humility we have lost is the recognition that Pauling and Szent-Gyorgyi (and Bronowski) were GIANTS! And now, we stand deeply impoverished by their loss and wonder whether life has any meaning at all, or shall we just all toss it in, and let a bunch of robots, without a political agenda, proceed on their own path toward discovery. A path which clearly does not lead to a world which combines 'Science and Human Values', the title of one of Bronowski's books, but leads to knowledge of, by and for a world of robots. Such a vision feels to me more out of the tradition of Asimov or Bradbury, than that of Pauling, Szent-Gyorgyi or Bronowski. Though all five of these authors manifest some kind of genius, the latter three were clearly of this earth. The former two spent a good deal of their time on what I think was a lesser planet, one of their own creation. Bert Gold San Francisco Dima Klenchin (klenchin@macc.wisc.edu) wrote: : In article , : Bert Gold wrote: : Newsgroups: : >bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bi : >onet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience : >,sci.med,sci.research.careers,talk.politics,talk.politics.medicine : >My hope in writing this is that our children will not suffer : >because of the lack of wisdom of their leaders in : >making decision about what to and what not to study. : Judging by the number of newsgroups crossposted, Bert Gold fully qualifies : as spammer... : :-)) Just kidding : Seriously: Dr. Gold has obviously very strong opinion on the subject. The : Q: is it really confirmed by research, or the current state of affair : is simply that kilogram amounts of ascorbic acid don't hurt (apparently)? : - Dima From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!news.uscyber.com!usenet From: dcesh3 Newsgroups: bionet.molbio.proteins Subject: Seven proteins envolving the Ebola Virus Date: 7 May 1996 15:25:31 GMT Organization: US Cyber Lines: 8 Message-ID: <4mnq1b$ppf@newsie.uscyber.com> NNTP-Posting-Host: ppp006-waus.wis.com My name is Kelly, I'm doing a Term paper on the Ebola Virus. I'd Appreciate any possible insight that people may have on the topic of the seven proteins making up the Ebola Virus. Only three proteins are known right now. Whould anyone know anything about those three and if so write to me. The help would greatly be appriciated. From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!news.uscyber.com!usenet From: dcesh3 Newsgroups: bionet.molbio.proteins Subject: Seven proteins envolving the Ebola Virus Date: 7 May 1996 15:25:23 GMT Organization: US Cyber Lines: 8 Message-ID: <4mnq13$ppa@newsie.uscyber.com> NNTP-Posting-Host: ppp006-waus.wis.com My name is Kelly, I'm doing a Term paper on the Ebola Virus. I'd Appreciate any possible insight that people may have on the topic of the seven proteins making up the Ebola Virus. Only three proteins are known right now. Whould anyone know anything about those three and if so write to me. The help would greatly be appriciated. From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!agate!cgl!itssrv1.ucsf.edu!itsa.ucsf.edu!bgold From: bgold@itsa.ucsf.edu (Bert Gold) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Followup-To: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Date: 7 May 1996 19:25:43 GMT Organization: UCSF, ITS Lines: 69 Message-ID: <4mo83n$103v@itssrv1.ucsf.edu> References: <318E9B47.285B@indirect.com> NNTP-Posting-Host: itsa.ucsf.edu X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.general:21527 bionet.biology.cardiovascular:946 bionet.cellbiol:4626 bionet.glycosci:665 bionet.biophysics:1944 bionet.molbio.ageing:2694 bionet.molbio.proteins:7818 bionet.neuroscience:13987 sci.med:122643 sci.research.careers:10149 talk.politics.medicine:51242 I agree with Mr. Wagner when he suggests that Mr. Kuslich has not properly sorted the wheat from the chaff in the scientific literature. In addition, it is precisely the point that science has become overly politicized which I was trying to make in writing my post. In fact, regular readers of my essays will remember that the Woods Hole meeting I proposed specifically excluded media and was wholly aimed at gathering a fair and honest consensus from scientists before presenting the results to the public. However, the scientific ruling elite have chosen not to follow this wise course, preferring noisy public debate and namecalling to reasoned, well thought-out discourse. Our leaders need to rethink their positions. Bert Gold San Francisco Path: itssrv1.ucsf.edu!cgl!sgigate.sgi.com!nntp.coast.net!news.kei.com!newsfeed.internetmci.com!uwm.edu!math.ohio-state.edu!pacific.mps.ohio-state.edu!pacific.mps.ohio-state.edu!not-for-mail From: wagner@pacific.mps.ohio-state.edu (Lukas Wagner) Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: 7 May 1996 12:06:34 -0400 Organization: Ohio State University Physics Dept. John E. Kuslich wrote: >Just look at research on "Cold Fusion", "Global Warming", and "The Ozone >Hole" for further examples of the poor quality of some recent so-called >scientific research. > The ozone hole is quite real. You might look up articles by Sherwood Rowland written in the last 10 years if you don't believe me, or if you have qualms about the quality of the research. The evidence on global warming is mixed to the best of my knowledge. I believe that the basis for the conclusions is careful work on ice cores. Why you put either of these in the same category as cold fusion is beyond me; perhaps you can cite review articles in real journals that indicate the poor quality of the science? >Chaos theory eliminated any hope of long term weather prediction, and >yet we still have "researchers" at major universities doing computer >simulation of weather. > Generously, this sentence suggests that you haven't read the Journal of Atmospheric Research recently. If getting to a library is troublesome, catch NCAR's web site: http://www.ncar.ucar.edu/ for some idea of what people who study weather actually do. The Lyapunov spectrum of weather models depends on the number of modes one includes. The number of days over which it's feasible to forecast weather depends on the largest Lyapunov exponent. You might look at J. Curry, Commun. Math. Phys. 60, 193 (1978). I hope that what you do for a living is less half-assed than your post. Science has some real problems; excessive politicization is certainly one of them. What you seem to cite as symptoms aren't. -- -Lukas Wagner wagner@pacific.mps.ohio-state.edu From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!newsfeed.pitt.edu!bb3.andrew.cmu.edu!andrew.cmu.edu!mm6n+ From: Marcella Madrid Newsgroups: bionet.molbio.proteins Subject: Sequence Analysis workshops offered at the PSC Date: Tue, 7 May 1996 15:06:50 -0400 Organization: Pittsburgh Supercomputing Center, Carnegie Mellon, Pittsburgh, PA Lines: 122 Message-ID: <8lXtx_y00YUpIca4Uv@andrew.cmu.edu> NNTP-Posting-Host: po8.andrew.cmu.edu NUCLEIC ACID AND PROTEIN SEQUENCE ANALYSIS WORKSHOPS FOR BIOMEDICAL RESEARCHERS PITTSBURGH SUPERCOMPUTING CENTER Biomedical Workshops offered by the Pittsburgh Supercomputing Center typically consist of theoretical lectures taught by leaders in the respective scientific discipline, and extensive hands-on computer sessions. During the computer sessions, participants are able to work on the examples provided or on their own experimental data. Attendance is limited to 20 participants to allow one-on-one instruction and encourage scientific interactions and discussions. Researchers nationwide are invited to apply. For additional information, please refer to http://www.psc.edu/biomed/workshops.html CONTACT INFORMATION: Nancy Blankenstein, Biomedical Program Assistant, (412)268-4960, blankens@psc.edu The following two Sequence Analysis workshops will be offered this summer: *********NUCLEIC ACID AND PROTEIN SEQUENCE ANALYSIS: June 9-14.************** Emphasis will be on alignment of and pattern extraction from multiple sequences. Topics to be discussed include Comparing and aligning sequences Identifying informative patterns in a set of sequences Using extracted informative patterns to identify related sequences. Leaders are: Dr. Gary Churchill, Cornell University; Dr. Michael Gribskov, San Diego Supercomputer Center and Dr. Hugh B. Nicholas Jr., Pittsburgh Supercomputing Center. Financial Information: A limited number of grants to cover travel and/or hotel accommodations are available for U.S. academic participants. ALL PARTICIPANTS ARE REQUIRED TO PAY A $135 REGISTRATION FEE UPON ACCEPTANCE INTO THE WORKSHOP. Complimentary breakfast and lunches will be provided. APPLICATION DEADLINE: May 17, 1996. *****ADVANCED NUCLEIC ACID AND PROTEIN SEQUENCE ANALYSIS: August 25-28.****** Open to researchers who have previously attended one of the PSC's annual "Nucleic Acid and Protein Sequence Analysis" workshops or who have appreciable experience with computerized sequence analysis. The workshop will build on previous experience to teach techniques for analyzing families and superfamilies of genes and proteins. Leaders are: Dr Stephen H. Bryant, National Center for Biotechnology Information, National Library of Medicine Dr. Michael Gribskov, San Diego Supercomputer Center and Dr. Hugh B. Nicholas Jr., Pittsburgh Supercomputing Center. Financial Information : Hotel accommodations during the workshop for researchers affiliated with U.S. academic institutions will be paid by our NIH grant. Complimentary breakfast and lunches will also be provided. There is NO REGISTRATION FEE for this workshop. All other costs incurred in attending (travel, other meals, etc.) are the responsibility of the individual participant. APPLICATION DEADLINE: July 10, 1996. ********** PITTSBURGH SUPERCOMPUTING CENTER WORKSHOPS FOR BIOMEDICAL RESEARCHERS APPLICATION Workshop I am interested in attending:_______________________________________ Name: ______________________________________________________________ Affiliation: ______________________________________________________________ Address: ______________________________________________________________ (Business) ______________________________________________________________ ______________________________________________________________ (Home) ______________________________________________________________ Telephone: ____________________________ ____________________________ (Business) (Home) *Social Security Number: _______-_____-_______ Citizenship:_________________ Electronic Mail Address:_____________________________________________________ Status: ___Graduate ___Post-doctoral Fellow ___Faculty ___Other (specify) Please indicate specifically any special housing, transportation or dietary arrangements you will need: __________________________________________ How did you learn about this workshop:_______________________________________ REQUIREMENTS: Applicants must submit a completed application form and a cover letter. The letter should describe, in one or two paragraphs, the sequence analysis problems encountered in your research, and how participating in the workshop will enhance this research. Please include a brief statement describing your level of experience with computers. Faculty members, staff and post-docs should provide a curriculum vitae. Graduate students must have a letter of recommendation from a faculty member. Please return all application materials to: Biomedical Workshop Applications Committee Pittsburgh Supercomputing Center 4400 Fifth Avenue, Suite 230C Pittsburgh, PA 15213 Direct inquiries to: Nancy Blankenstein, blankens@psc.edu or 412/268-4960. *Disclosure of Social Security Number is voluntary. PSC does not discriminate on the basis of race, color, religion, sex, age, creed, national or ethnic origin, or handicap. From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!usenet From: Ilia Ouspenski Newsgroups: bionet.molbio.proteins Subject: Re: Aggregation of His-tagged proteins Date: 7 May 1996 23:48:43 GMT Organization: Baylor College of Medicine Lines: 5 Message-ID: <4mongr$9ip@gazette.bcm.tmc.edu> References: <4mdufk$37u@freenet-news.carleton.ca> NNTP-Posting-Host: ao.cellb.bcm.tmc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: at332@FreeNet.Carleton.CA We had a similar problem. In our hands, the protein seemed to be more soluble at very high (>9) or very low (<4) pH. We did not pursue it further, but would be very interested to know if there is a way around it. From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!globe.indirect.com!usenet From: "John E. Kuslich" Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: Tue, 07 May 1996 15:12:42 -0600 Organization: CRAK Software Lines: 107 Message-ID: <318FBCCA.355B@indirect.com> References: <318E9B47.285B@indirect.com> <4mo6if$9vs@frazier.uoknor.edu> NNTP-Posting-Host: s141.phxslip4.indirect.com Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0 (Win95; I) CC: news Xref: biosci bionet.general:21529 bionet.biology.cardiovascular:947 bionet.cellbiol:4627 bionet.glycosci:667 bionet.biophysics:1945 bionet.molbio.ageing:2695 bionet.molbio.proteins:7821 bionet.neuroscience:13989 sci.med:122659 sci.research.careers:10150 talk.politics.medicine:51247 Scott Russell wrote: > > In article <318E9B47.285B@indirect.com>, > "John E. Kuslich" wrote: > >Bert Gold wrote: > >> <<<>> > > > >> And yet, this week, fully 18 years after the memorable talk by > >> Szent-Gyorgyi which I just described, the Vitamin C RDA for men > >> has been adjusted three-fold upward. > >> > >> It makes me wonder at the brazen inefficiency of the research > >> 'establishment'... > >> > >> > >> Bert Gold > >> San Francisco > >> > >> References > > > ><<> > > > >Just look at research on "Cold Fusion", "Global Warming", and "The Ozone > >Hole" for further examples of the poor quality of some recent so-called > >scientific research. > > > >Chaos theory eliminated any hope of long term weather prediction, and > >yet we still have "researchers" at major universities doing computer > >simulation of weather. > > > >Modern science has gone "politically correct"; look at the reaction to > >that book entitled "The Bell Curve". It was widely renounced by many > >so-called scientists who never took the time to read it!!! > > > >Oh well... > > > >John E. Kuslich > > Now wait a minute. > > (1) None of the topics in the first paragraph have been eliminated as > possibilities, the first two are just not supported by sufficient evidence to > warrant the conclusions that were made. I think you effectively illustrate attitudes that lead us to accept conclusions of and act upon "junk " science. I believe that if you examine the case for all three of the topics mentioned, you will be led to the inescapable conclusion that "Junk" science is alive and well. Some of these nutty ideas only gain popular acceptance because of the tendency for the liberal media to publicize them - they fit well into the environmentalist agenda. "Possibilities" don't necessarily merit action which will cause serious economic dislocations. The "Carbon Tax" comes to mind. > > (2) That weather is ultimately unpredictable does not mean that extending > forecasts is not useful. Chaos theory indicated that predicting the behavior > of self-iterating systems using incomplete data can be expected to be > unsuccessful over the course of a certain amount of time. > You misstate my point. Chaos theory indicates quite convincingly that there are severe limits on the ability of computers to make long term weather predictions because we simply 1) can never model the physical weather interactions adequately, 2) we can never have enough knowledge of initial condidtions to put into our model to get long term results. We might as well put research money into perpetual motion. That was my point. > (3) "The Bell Curve" received almost unanimous negative reviews from > scientists because measuring intelligence was treated as if: (a) we knew what > intelligence was; (b) we could measure it reproducibly and accurately; and (c) > that it is possible to any of this in a culturally neutral manner. Most were > unimpressed by the numerous charts and graphs because underlying them, the > science was weak and the "parameters" were meaningless. If the IQ test > measures anything, it measures the ability of people to take IQ tests, and it > is not even entirely reliable for that. Given the absence of compelling data, > the book has a pernicious effect on society and its conclusions ignore > too many societal effects on those measured. The use of "science" to achieve > social ends based on race should doomed to fail because, this approach like > Nazi "science," is not based on a single uncontested fact. Gees. The point I was making is perfectly illustrated by your paragraph above. Real research and real science do not question the political implications of the truth. I don't know if the conclusions of this book are good ones or not. The point is, this kind of subject matter is avoided by science for politcal reasons. That can hardly be a method that will lead us to the truth!! It is like the Vatican criticizing fertility research based on it's position on birth control. If you don't do birth control research, you won't arrive at the truth as to which methods are effective. Similarly, if you cannot state certain hypothyses and examine the implications of such an hypothyses because they are not politically correct, then you are not doing science. Nazi science is as bad as un-Nazi science, or Vatican science or Zionist science or Environmentalist science. If it is limited in scope or subject for political reasons, it is not science. That was my point. > -- John E. Kuslich WPcrak for Wordperfect johnk@indirect.com WDcrak for MS Word Password Recovery EXcrak for MS Excel 602 863 9274 voice QPcrak for Quattro Pro 602 548 1993 fax LOcrak for Lotus 123 Password Removal QWcrak for Quicken Our WEB Site http://www.indirect.com/www/johnk/ >>>>>>>>You Hack'em, We CRAK'em<<<<<<<<<<<<< From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer2.itd.umich.edu!uunet!in1.uu.net!mail.llu.edu!usenet From: bLangridge@ccmail.llu.edu (Bill Langridge) Newsgroups: bionet.molbio.proteins Subject: Soybeans Date: 8 May 1996 04:43:29 GMT Organization: Loma Linda University Lines: 13 Message-ID: <4mp8ph$iia@mail.llu.edu> NNTP-Posting-Host: 151.112.27.53 Mime-Version: 1.0 Content-Type: Text/Plain; charset=ISO-8859-1 X-Newsreader: WinVN 0.99.5 Can anyone tell me what the present yield of soybeans is in the U.S.? I am trying to determine what the yield of recombinant protein might be in soybean as compared to what is presently able to be synthesized in transgenic potato. Are there any good (high expression) soybean promoters available? Thanks very much for your time, patience and help! Bill Langridge Department of Biochemistry Center for Molecular Biology Loma Linda Univesity, L.L. CA.92350 blangridge@ccmail.llu.edu From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!EU.net!newsfeed.internetmci.com!in2.uu.net!news3.ottawa.istar.net!istar.net!news1.vancouver.istar.net!news.vancouver.istar.net!west.news.istar.net!van-bc!unixg.ubc.ca!hayci.generes.ca!user From: ky@ulam.generes.ca (rona graham) Newsgroups: bionet.molbio.proteins Subject: Western Control Date: Tue, 07 May 1996 08:28:37 -0800 Organization: ubc Lines: 3 Message-ID: NNTP-Posting-Host: hayci.generes.ca I'm looking for an antibody that i can use on a western to control for loading, degradation and transfer of a total protein prep (cytosolic fraction). Something like actin is to a northern. any ideas? From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!EU.net!sun4nl!surfnet.nl!swsbe6.switch.ch!swidir.switch.ch!in2p3.fr!univ-lyon1.fr!pasteur.fr!jussieu.fr!citi2.fr!camoin.cochin.inserm.fr!user From: brydon@icgm.cochin.inserm.fr (Lena Brydon) Newsgroups: bionet.molbio.proteins Subject: Review Antibody Production Date: Tue, 07 May 1996 17:52:06 +0200 Organization: icgm cnrs upr415 Lines: 24 Message-ID: NNTP-Posting-Host: camoin.cochin.inserm.fr X-Newsreader: Yet Another NewsWatcher F2.0 Dear Netters, I am compiling a review of existing and proposed methods for the production and utilisation of monoclonal and polyclonal antibodies. I would be very grateful for any information/ideas you have regarding this subject area. The results of this inquiry will be sent to this news group later. If anyone has any papers recently published or in Press regarding this subject and are willing to send me re-prints, I will of course quote these authors in the review ( if so desired.) Thanks to all those who have already responded! (Especially Keld Sorensen) The following is my snail mail address for anyone else who would like to send me ideas/info./re-prints: Lena Brydon, ICGM, CNRS UPR415, 22 rue Mechain-75014 PARIS, FRANCE. Fax. (331) 40.51.72.10 From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!enews.sgi.com!sgigate.sgi.com!news.msfc.nasa.gov!info.uah.edu!maze.dpo.uab.edu!usenet From: "Richard J. Dudley" Newsgroups: bionet.molbio.proteins Subject: Re: Protease activity determination Date: 7 May 1996 14:52:51 GMT Organization: nrc Lines: 15 Message-ID: <4mno43$urh@maze.dpo.uab.edu> References: <318F21D4.56F4@nibh.go.jp> NNTP-Posting-Host: 138.26.50.136 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; PPC) To: benoit@nibh.go.jp X-URL: news:318F21D4.56F4@nibh.go.jp No kit that I know of will detect all proteases, but I think Pierce ans Sigma sell kits that are pretty general. I have used FITC-labelled BSA and LEU-methylcoumarinamide in the past, but they are fluorometric. Azocoll or azocasein is colorometric. The fluoroscent assays are simpler. There are also some peptidase substrates that use p-nitroaniline, which absorbs at 410 nm as a free product. I don't have a lot if info in fron of me, but if you e-mail me and remind me, I can track my folder down. --- --- --- Richard J. Dudley Neurobiology Research Center University of Alabama School of Medicine From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!enews.sgi.com!sgigate.sgi.com!news.msfc.nasa.gov!info.uah.edu!maze.dpo.uab.edu!usenet From: "Richard J. Dudley" Newsgroups: bionet.molbio.proteins Subject: Re: Where can I find a database of peptidases? Date: 7 May 1996 14:48:31 GMT Organization: nrc Lines: 10 Message-ID: <4mnnrv$urh@maze.dpo.uab.edu> References: NNTP-Posting-Host: 138.26.50.136 Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; PPC) To: oravaxcm@world.std.com X-URL: news:Dr0pMs.36u@world.std.com Try starting at Pedro's tools--there are several enzyme databases you can search. The address is: http://www.public.iastate.edu/~pedro/research_tools.html --- --- --- Richard J. Dudley Neurobiology Research Center University of Alabama School of Medicine From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!nntp.coast.net!torn!resunix.ri.sickkids.on.ca!news From: Randall Willis Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: Q: Properties of Phospho-cellulose (P11, Whatman) Date: 7 May 1996 15:06:11 GMT Organization: The Hospital for Sick Children Lines: 7 Message-ID: <4mnot3$qfc@resunix.ri.sickkids.on.ca> References: <4mjcme$6j8_001@biochem.wisc.edu> <4mksmq$9l2@resunix.ri.sickkids.on.ca> NNTP-Posting-Host: resunix.ri.sickkids.on.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) To: maga@vetbio.unizh.ch X-URL: news:maga-0705960922510001@130.60.120.11 Xref: biosci bionet.molbio.methds-reagnts:44166 bionet.molbio.proteins:7809 You're a better man than I, McGinty. I'm saving your posting and will try your system...thanks for the info. Randall C Willis From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!uwvax!newssinet!news.nc.u-tokyo.ac.jp!zion.phys.s.u-tokyo.ac.jp!news.tisn.ad.jp!ripspost.aist.go.jp!usenet From: Beno=?iso-8859-1?Q?=EEt VIARD ?= Newsgroups: bionet.molbio.proteins Subject: Protease activity determination Date: Tue, 07 May 1996 19:11:32 +0900 Organization: Agency of Industrial Science and Technology, Tsukuba Lines: 16 Message-ID: <318F21D4.56F4@nibh.go.jp> NNTP-Posting-Host: ripspom.aist.go.jp Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.01 (Macintosh; I; 68K) Hello Netters, I am looking for a protease activity determination kit or a protocol for protease determination in culture medium. Maybe Azocoll ? Could someone give me advice ? Sincerely, Benoit VIARD, Postdoctoral Researcher Biochemical Engineering Laboratory National Institute of Bioscience and Human-Technology 1-1, Higashi, Tsukuba Ibaraki 305, JAPAN Tel : 81 298 54 6087 Fax : 81 298 54 6080 From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!ipc.pku.edu.cn!ldw From: ldw@ipc.pku.edu.cn (ldw) Newsgroups: bionet.molbio.proteins Subject: ANNOUNCE- New Bioinformatics server in China (fwd) Date: 7 May 1996 05:47:27 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 55 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear Collages, We are pleased to announce that the first bioinformatics server in China has been setup. It is now maintained by Molecular Design Lab, Institute of Physical Chemistry, Peking University, Beijing, China. At this infant stage, it has only mirrored protein related databases, other databases of molecular biology will soon be mirrored as requested. Under the direction of Prof. Luhua Lai, all the mirror work and database interconnections are implenmented by Mr. Dawei Lin. The Home Pages were designed by Mr. Renxiao Wang and Mr. Dawei Lin. The work is supported by the Chinese National Commision of Science and Technology. At present, we are one of three official mirror sites of PDB(Protein Data Bank) including all the ftp directories and WWW browsing. We are one of five official SCOP mirror sites. We also mirrored SWISSPROT, PIR, PROSITE, BLOCKS, but only ftp services can be provided at present. The PDB and SCOP mirror sites have been crosslinked for the convenient use of local people. Also a public molecular graphics software-Rasmol is attached to the databases. Users in China and nearby coutries or areas may find this server helpful. We would like to thank all the people who give us help and valuable suggestions and specail thanks to Prof. Joel L. Sussman, Prof. Enrique Abola, Prof. Jaime Prilusky, Dr. Dave Stampf, Dr. Nancy O. Manning for the help in setting up PDB mirror site, to Dr. Steven Brenner and Prof. Tim Hubbard for the help in setting up SCOP mirror site, to Prof. Amos Bairoch for the help in setting up SwissProt mirror site. The useful URLs related to the bioinformatics server are: The first China bioinformatics server home page: http://www.ipc.pku.edu.cn/mirror/mirror.html The home page of Institute of Physical Chemistry, Peking University: http://www.ipc.pku.edu.cn/ The PDB mirror site page: http://www.ipc.pku.edu.cn/npdb/index.html The SCOP mirror site page: http://www.ipc.pku.edu.cn/scop/ The anonymous ftp server name of IPC is: ftp.ipc.pku.edu.cn Suggestions and encouragements are welcome at lai@ipc.pku.edu.cn ----------< *** Dawei LIN *** >---------------- Ph.D student of Molecular Design Lab Institute of Physical chemistry Peking University Beijing 100871 CHINA Phone: 86-10-2751490 Fax: 86-10-2751725 Email: ldw@pschnetware.pku.edu.cn ldw@ipc.pku.edu.cn URL: http://162.105.177.12/moldes/ldw/ldw.htm ------------------------------------------------ From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!newsfeed.internetmci.com!info.ucla.edu!psgrain!quagga.ru.ac.za!inet.up.ac.za!crimson!fourie From: fourie@crimson (Fourie Joubert) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Human DHFR clones? Date: 7 May 1996 10:53:27 GMT Organization: University of Pretoria Lines: 19 Message-ID: <4mna37$15j4@inet.up.ac.za> Reply-To: fourie@scientia.up.ac.za NNTP-Posting-Host: crimson.up.ac.za X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.molbio.methds-reagnts:44158 bionet.molbio.proteins:7806 Hi I am trying to obtain clones of human DHFR in any vector. If anyone would be willing to share, please let me know. We are working on the malaria form of the enzyme and would like to do some comparisons... Any help would be sincerely appreciated! -------------- Fourie Joubert Department of Biochemistry University of Pretoria fourie@scientia.up.ac.za http://suntiger.ee.up.ac.za/biodocs/home.html +27-12-420-2011 From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!daresbury!yama.mcc.ac.uk!sunsite.doc.ic.ac.uk!nntp0.brunel.ac.uk!strath-cs!queens-belfast.ac.uk!bcg0197 From: bcg0197@queens-belfast.ac.uk (Andrew Wallace) Newsgroups: bionet.molbio.proteins Subject: Postgraduate studentship available Message-ID: <1996May7.105718.6294@queens-belfast.ac.uk> Date: 7 May 96 10:57:18 GMT References: <1996May7.104523.6292@queens-belfast.ac.uk> X-From: bcg0197@queens-belfast.ac.uk (Andrew Wallace) X-Newsgroups: bionet.molecules.repertoires,bionet.jobs Followup-To: bionet.molbio.proteins,bionet.molecules.repertoires,bionet.jobs X-Date: 7 May 96 10:45:23 GMT X-Organization: Queen's University of Belfast Lines: 35 X-Xref: queens-belfast.ac.uk bionet.molecules.repertoires:69 bionet.jobs:3740 A Postgraduate Research Studentship is available in my laboratory commencing in October 1996. Two projects are currently available to choose from: either: use of combinatorial libraries of mechanism-specific active site-directed protease inhibitors to identify novel proteases involved in the regulation of apoptosis. or: generation of catalytic antibodies with proteolytic activity by selection from a phage-displayed antibody library using novel synthetic transition-state analogues. Applications are invited from UK and EU candidates who have or expect to receive a relevant First or Upper Second Class Honours degree or equivalent qualification, or from qualified non-EU international students who can obtain their own funding. For UK and EU canditates, the studentship pays tuition fees, laboratory costs and a living expenses contribution of about 1000 pounds sterling per quarter year. Further details and application forms are available from: The Director, School of Biology and Biochemistry The Queen's University of Belfast 97 Lisburn Road Tel. +44-1232-335786 Belfast BT9 7BL Fax +44-1232-236505 Northern Ireland (UK) sobb.office@queens-belfast.ac.uk All applications must be received by the Admissions Office, The Queen's University of Belfast no later than 15 May 1996 -- ============================================================ Andrew Wallace, Ph.D. School of Biology and Biochemistry From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!nntp.coast.net!howland.reston.ans.net!math.ohio-state.edu!jussieu.fr!univ-lyon1.fr!in2p3.fr!swidir.switch.ch!scsing.switch.ch!rzunews.unizh.ch!NewsWatcher!user From: maga@vetbio.unizh.ch (Giovanni Maga) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: Q: Properties of Phospho-cellulose (P11, Whatman) Date: Tue, 07 May 1996 09:22:51 -0500 Organization: University of Zurich Irchel- Biochemistry Lines: 35 Message-ID: References: <4mjcme$6j8_001@biochem.wisc.edu> <4mksmq$9l2@resunix.ri.sickkids.on.ca> NNTP-Posting-Host: 130.60.120.11 Xref: biosci bionet.molbio.methds-reagnts:44151 bionet.molbio.proteins:7804 In article <4mksmq$9l2@resunix.ri.sickkids.on.ca>, Randall Willis wrote: > Dima, > > Unfortunately, what Whatman says is true. I have worked with P11 off > and on for over 8 years and have found it to be the most intractable > resin on the market. well, I used PC for a comparable amount of years for purification of DNA polymerases from eukaryotic cells and I had very little problems with it. Let's say that on a PC you should not expect to get high resolution, but it's extremely useful as a first column for fractionation of crude extracts if you are looking for a DNA binding protein. I used it in either 20 mM KPO4, pH 7.0, eluting it with a single step at 350 mM KPO4, or in 50 mM TrisHCl, pH 7.5, eluting with a NaCl gradient. Typically, less then 30% of the total proteins will bind, but more then 70% of the total DNA polymerase activity is retained. After using, I regenerate it by washing with 1 M KPO4, pH 7.0 (in the first protocol) or 2M NaCl in 50 mM TrisHCl pH 8.5 (in the second case). Storage is in 20 mM KPO4, pH 7.0, 0.01% Sodium Azide. I found reproducible patterns of elution up to 1 year of storage at 4°C. I strongly recommend, anyway, to check the pH after prolonged storage, since PC is quite sensitive to pH changes (indeed, you can elute it also with a pH gradient, even if the resolution is not very good). If you are using a new batch (i.e. prepared from fresh powder), after the treatment suggested by the booklet (prolonged washing with high-/low- pH) I would run a first column with a crude extract with step elution in high salt, in order to remove completely eventual unspecific inhibitors. Actually, the most used it is, the best yeld you get. Another tip...use a column with a diameter of about 1/3 or 1/2 of the lenght. Fat, short columns are better than long, thin ones with PC, especially if the volume is large. The opposite is true if you use fast flow, high resolution cation exchangers like S-Sepharose or SP-Sepharose (that are also stronger, so keep it in mind if your protein has a highly basic pI). Hope it helps. From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!EU.net!newsfeed.internetmci.com!uwm.edu!spool.mu.edu!munnari.OZ.AU!news.ecn.uoknor.edu!news.uoknor.edu!botrusse From: srussell@ou.edu (Scott Russell) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: Tue, 07 May 96 18:54:37 GMT Organization: University of Oklahoma Lines: 69 Message-ID: <4mo6if$9vs@frazier.uoknor.edu> References: <318E9B47.285B@indirect.com> NNTP-Posting-Host: botrusse.bio.uoknor.edu X-Newsreader: News Xpress Version 1.0 Beta #4 Xref: biosci bionet.general:21524 bionet.biology.cardiovascular:945 bionet.cellbiol:4624 bionet.glycosci:664 bionet.biophysics:1943 bionet.molbio.ageing:2693 bionet.molbio.proteins:7816 bionet.neuroscience:13986 sci.med:122640 sci.research.careers:10148 talk.politics.medicine:51241 In article <318E9B47.285B@indirect.com>, "John E. Kuslich" wrote: >Bert Gold wrote: >> <<<>> > >> And yet, this week, fully 18 years after the memorable talk by >> Szent-Gyorgyi which I just described, the Vitamin C RDA for men >> has been adjusted three-fold upward. >> >> It makes me wonder at the brazen inefficiency of the research >> 'establishment'... >> >> >> Bert Gold >> San Francisco >> >> References > ><<> > >Just look at research on "Cold Fusion", "Global Warming", and "The Ozone >Hole" for further examples of the poor quality of some recent so-called >scientific research. > >Chaos theory eliminated any hope of long term weather prediction, and >yet we still have "researchers" at major universities doing computer >simulation of weather. > >Modern science has gone "politically correct"; look at the reaction to >that book entitled "The Bell Curve". It was widely renounced by many >so-called scientists who never took the time to read it!!! > >Oh well... > >John E. Kuslich Now wait a minute. (1) None of the topics in the first paragraph have been eliminated as possibilities, the first two are just not supported by sufficient evidence to warrant the conclusions that were made. (2) That weather is ultimately unpredictable does not mean that extending forecasts is not useful. Chaos theory indicated that predicting the behavior of self-iterating systems using incomplete data can be expected to be unsuccessful over the course of a certain amount of time. (3) "The Bell Curve" received almost unanimous negative reviews from scientists because measuring intelligence was treated as if: (a) we knew what intelligence was; (b) we could measure it reproducibly and accurately; and (c) that it is possible to any of this in a culturally neutral manner. Most were unimpressed by the numerous charts and graphs because underlying them, the science was weak and the "parameters" were meaningless. If the IQ test measures anything, it measures the ability of people to take IQ tests, and it is not even entirely reliable for that. Given the absence of compelling data, the book has a pernicious effect on society and its conclusions ignore too many societal effects on those measured. The use of "science" to achieve social ends based on race should doomed to fail because, this approach like Nazi "science," is not based on a single uncontested fact. ========================================================================= Scott D. Russell Internet: srussell@ou.edu Dept of Botany & Microbiology ->http://www.ou.edu/cas/botany-micro/ & Noble Electron Microscopy Lab ->http://www.ou.edu/research/electron/ University of Oklahoma, Norman OK Phone: 1-405-325-6234 73019-0245 USA FAX: 1-405-325-7619 =========================================================================  From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!agate!nature.berkeley.edu!lhom From: lhom@nature.berkeley.edu (Louis Hom) Newsgroups: bionet.molbio.proteins,bionet.molbio.methds-reagnts,sci.chem Subject: Small molecules & nitrocellulose Date: 7 May 1996 17:10:37 GMT Organization: University of California, Berkeley Lines: 10 Message-ID: <4mo06d$rae@agate.berkeley.edu> NNTP-Posting-Host: nature.berkeley.edu Xref: biosci bionet.molbio.proteins:7815 bionet.molbio.methds-reagnts:44175 sci.chem:55599 I'm looking to remove proteins from a small volume of a complex biological mixture (too small for ultrafiltration, IMHO) and I'm thinking maybe I could wet a piece of nitrocellulose and stick it into my sample to do the job. But I'm wondering, what might the nitrocellulose remove besides the intended proteins? -- _______________________________________________________________________________ Lou Hom >K '93 "Is that a deliberate act of provocation lhom@nature.berkeley.edu or just a parochial New Yorkism?" http://www.ocf.berkeley.edu/~lhom From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!ns1.faseb.org!lamarck.sura.net!newsfeed.internetmci.com!uwm.edu!vixen.cso.uiuc.edu!news.uoregon.edu!raffles.technet.sg!nf1.iij.ad.jp!news.iij.ad.jp!wnoc-tyo-news!wnoc-sfc-news!wnoc-kyo-news!kuis-news!s2000!NewsWatcher!user From: nakai@imcb.osaka-u.ac.jp (Kenta Nakai) Newsgroups: bionet.molbio.proteins Subject: Re: Signal peptide sequence Date: Mon, 06 May 1996 13:39:35 +0900 Organization: Inst. Mol. Cell. Biol., Osaka U. Lines: 17 Distribution: world Message-ID: References: <4mc0b7$3oh@woodstock.socs.uts.EDU.AU> NNTP-Posting-Host: 133.1.132.91 In article <4mc0b7$3oh@woodstock.socs.uts.EDU.AU>, Slavica Masina wrote: > Does anybody know any methods, preferably recent, for the identification > of protein secretory sequences and for prediciting the site of cleavage > between a signal sequence and the mature exported protein. I currently > have the paper of G. von Heljne (1986) Nucleic acids research, 14(11), > but I would prefer computer program that performs the analysis. Can > anyone help? Try the PSORT WWW server (http://psort.nibb.ac.jp). It implements von Heijne's method for the prediction of signal peptide cleavage site and lots more for the prediction of subcellular localization site. Kenta Nakai IMCB, Osaka University From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!newsfeed.sunet.se!news01.sunet.se!sunic!news99.sunet.se!umdac!news From: Michael Daws Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins,bionet.molecules.peptides,bionet.cellbio,bionet.immunology Subject: Membrane-impermeable Reducing Agents Date: 6 May 1996 12:00:55 GMT Organization: University of Umea, Sweden Lines: 10 Message-ID: <4mkpln$oio@studium.student.umu.se> NNTP-Posting-Host: clsmac.ucmp.umu.se Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; PPC) X-URL: news:bionet.molbio.methds-reagnts Xref: biosci bionet.molbio.methds-reagnts:44202 bionet.molbio.proteins:7828 bionet.molecules.peptides:308 bionet.immunology:8757 I am looking to reduce disulphide bonds outside of cells without affecting internal disulphides. Is there a reducing agent I can use which is membrane impermeable. Alternatively is there an oxidising agent that is membrane impermeable? Thanks Mike Daws From owner-proteins@net.bio.net Mon May 06 23:00:00 1996 Path: biosci!IRIS.LSC.PKU.EDU.CN!lisl From: lisl@IRIS.LSC.PKU.EDU.CN (Li Songlin) Newsgroups: bionet.molbio.proteins Subject: Ebola Virus Date: 7 May 1996 21:52:57 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <9605081950.AA26017@IRIS.lsc.pku.edu.cn> NNTP-Posting-Host: net.bio.net In Message-Id: <4mnq1b$ppf@newsie.uscyber.com>, Kelly wrote, >I'm doing a Term paper on the Ebola Virus. I'd Appreciate any possible I believe that Ebola Virus is a subject for three dimmentional reconstruction. I do not remmenber who are working on it. Prof. Guangying Lu might give you some help, I believe. Songlin Li From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!newsfeed.internetmci.com!news.dacom.co.kr!news.kreonet.re.kr!usenet.kornet.nm.kr!snunews.snu.ac.kr!NewsWatcher!user From: juhnn@plaza.snu.ac.kr (YONG-SUNG JUHNN) Newsgroups: bionet.general,bionet.cellbiol,bionet.molbio.proteins,bionet.neuroscience Subject: receptors on COS cell membrane Date: Thu, 09 May 1996 14:23:45 +0900 Organization: DEPT OF BIOCHEMSITRY, SNU COLLEGE OF MEDICINE Lines: 10 Message-ID: NNTP-Posting-Host: 147.46.161.102 Xref: biosci bionet.general:21565 bionet.cellbiol:4647 bionet.molbio.proteins:7842 bionet.neuroscience:14016 Colleagues, I am planning to work on signal transduction with COS-1 cell line. I need to know what kinds of membrane receptors are endogeneously expressed on this cell line or COS-7 cells. I would be grateful, if you can tell me the expressed receptor types and the related reference by e.mail. My address is juhnn@plaza.snu.ac.kr Thank you. From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!galaxy.ucr.edu!library.ucla.edu!info.ucla.edu!psgrain!rainrgnews0!usenet From: chemica@bendnet.com (Dan Brose) Newsgroups: bionet.molbio.proteins,bionet.molbio.methds-reagnts,sci.chem Subject: Re: Small molecules & nitrocellulose Date: 9 May 1996 00:45:10 GMT Organization: Chemica Technologies, Inc. Lines: 18 Message-ID: <4mrf6m$npl@news0.rain.rg.net> References: <4mo06d$rae@agate.berkeley.edu> NNTP-Posting-Host: bend54.bendnet.com Mime-Version: 1.0 Content-Type: Text/Plain; charset=US-ASCII X-Newsreader: WinVN 0.99.7 Xref: biosci bionet.molbio.proteins:7840 bionet.molbio.methds-reagnts:44248 sci.chem:55689 In article <4mo06d$rae@agate.berkeley.edu>, lhom@nature.berkeley.edu says... > >I'm looking to remove proteins from a small volume of a complex biological >mixture (too small for ultrafiltration, IMHO) and I'm thinking maybe I >could wet a piece of nitrocellulose and stick it into my sample to do the >job. But I'm wondering, what might the nitrocellulose remove besides the >intended proteins? >-- >______________________________________________________________________________ _ >Lou Hom >K '93 "Is that a deliberate act of provocation >lhom@nature.berkeley.edu or just a parochial New Yorkism?" >http://www.ocf.berkeley.edu/~lhom Is your sample too small to use the small centrifuge vials with UF membranes within the centrifuge vial. I believe you can filter quite small samples by this technique. From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!galaxy.ucr.edu!library.ucla.edu!info.ucla.edu!agate!cgl!itssrv1.ucsf.edu!itsa.ucsf.edu!bgold From: bgold@itsa.ucsf.edu (Bert Gold) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Followup-To: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Date: 8 May 1996 20:30:55 GMT Organization: UCSF, ITS Lines: 45 Distribution: world Message-ID: <4mr09v$14q1@itssrv1.ucsf.edu> References: <4mqk1n$hah@cosmos.uthscsa.edu> NNTP-Posting-Host: itsa.ucsf.edu X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.general:21556 bionet.biology.cardiovascular:954 bionet.cellbiol:4644 bionet.biophysics:1955 bionet.molbio.ageing:2704 bionet.molbio.proteins:7839 bionet.neuroscience:14008 sci.med:122812 sci.research.careers:10168 talk.politics.medicine:51292 Dr. Deneke, If you bother to read Pauling, you will find that he not only advocated MEGAdose amounts of vitamin C, but was an ardent advocate of establishing a new RDA for ascorbate as well, based upon more controlled indicators than relief from scurvy; this is precisely the work which Levine et. al have now done, establishing a new RDA. The current study only establishes a new RDA for healthy, men non-smokers in their 20s. Dr. Pauling suggested that he thought anti-oxidant requirements would increase with increasing age. Levine et al. do not comment on this, or the RDA for women. Dr. Pauling suggested many other functions for Vitamin C than anti-oxidant effect: Few of these have been tested in the time which intervenes between his suggestions and now. It is notable that Dr. Levine did not cite Dr. Pauling or Dr. Szent-Gyorgyi in his PNAS publication. Further, in my telephone conversation with Dr. Levine last week, he admitted to me that funding at NIH for nutrition was "political". Bert Gold San Francisco : Pauling and Szent-Gyorgyi are my heros too, but neither a conspiracy or : scientific incompetence is necessary to explain delays in jumping to recommend : large doses of vitamin C. In fact chemistry, physiology etc. of ascorbic acid : has been under intense investigation over the last several decades. It is well : known that ascorbate, under some conditions acts a a pro-oxidant rather than an : antioxidant. As for all nutritional additives it is extremely difficult to : determine whether consumption of high levels of the pure product may cause : toxic effects which are neutralized by other components present in the natural : food sources. Do large doses of ascorbate protect, deplete, or block induction : of other antioxidants? I can probably find instances of all of these effects : in the literature of the past 5 years. In short, its not that simple. Evidence : for multiple benefits from increasing dietary intake of ascorbate and other : antioxidants by increased consumption of fruits and vegetables is very : convincing. The health benefits of ingesting large levels of purified : antioxidants of any kind are proving harder to analyse. Unfortunately we are : not yet to the point where we can usefully micro-manage our nutrition. Take : megadoses of vitamins if you chose, at your own risk, but many of us would : prefer to keep on eating our fruits and veggies. From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!biosci!not-for-mail From: Scott Le Grand Newsgroups: bionet.molbio.proteins,bionet.molec-model Subject: PDB3D Applet Date: 8 May 1996 12:59:06 -0700 Organization: University of California, Los Angeles Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <4moopp$1p1s@saba.info.ucla.edu> NNTP-Posting-Host: net.bio.net Xref: biosci bionet.molbio.proteins:7838 bionet.molec-model:918 Hi guys, I have a Java applet that allows one to view PDB files... it's sitting at http://www.mbi.ucla.edu/people/legrand/pdb.html. I designed it specifically to allow people to incorporate PDB structures into web pages. It allows one to rotate and scale reasonably large molecules in real-time, much faster than anything else I've seen so far. I'm looking for feedback on what else it needs... Of course, you need a Java-capable browser to see the thing... Scott Le Grand From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!nntp.uio.no!news.cais.net!news.mathworks.com!newsfeed.internetmci.com!in1.uu.net!pixie.tridom.com!usenet From: mp@eng.tridom.com (Mike Pitcher) Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: 8 May 1996 18:50:19 GMT Organization: AT&T Tridom Lines: 35 Message-ID: <4mqqdb$rfp@pixie.tridom.com> References: Reply-To: mp@eng.tridom.com NNTP-Posting-Host: yah.eng.tridom.com Xref: biosci bionet.general:21554 bionet.biology.cardiovascular:953 bionet.cellbiol:4641 bionet.glycosci:674 bionet.biophysics:1954 bionet.molbio.ageing:2703 bionet.molbio.proteins:7837 bionet.neuroscience:14005 sci.med:122791 sci.research.careers:10167 talk.politics.medicine:51290 I agree that the priorities may need a little adjusting and that the research funding process is not perfect. Since human health and well-being is such an important (and expensive) activity one would think that it would get top priority. Oh well. For what it's worth, let me share what I have found by trial and error. I live in Atlanta which is an area not particularly friendly to allergy sufferers. For 20 years, I would get respiratory problems two times a year (spring and fall) that would start with sore throat, coughing, sometimes leading to runny nose, watery eyes, occasionally sinus problems, etc. The Drs would inevitably prescribe an antibiotic and maybe cough syrup (if cough was a problem), Seldane, pseudephedrine hydrochloride (wasn't good for my BP), or whatever. Well, last year, I had enough of this crap. No fault of the Drs, but I decided to try vit C. Starting with 1000mg dose (500mg twice a day) last fall and now am down to 500mg once a day. The usual fall and spring resp. problems never appeared. I got a few sniffles, but *nothing* compared to what I usually get. No change in work conditions, same house, same car, etc. Only significant change (besides another yr older and a raise :-) ), is the vit C. Coincidence? I think not. Bottom line is, I think the RDA is totally out of whack. There needs to be a *lot* more research done with vitamins in general and focus on what it takes to make people function properly, rather than try to fix what's broken. Seems I remember an adage: An ounce of prevention is worth a pound of cure. How true. Mike Pitcher email: mp@eng.tridom.com From owner-proteins@net.bio.net Tue May 07 23:00:00 1996 Path: biosci!rutgers!gatech!news.cse.psu.edu!uwm.edu!vixen.cso.uiuc.edu!newsfeed.internetmci.com!swrinde!cs.utexas.edu!geraldo.cc.utexas.edu!netnews.uthscsa.edu!usenet From: S. Deneke Newsgroups: bionet.general,bionet.biology.cardiovascular,bionet.cellbiol,bionet.glycosci,bionet.biophysics,bionet.molbio.ageing,bionet.molbio.proteins,bionet.neuroscience,sci.med,sci.research.careers,talk.politics,talk.politics.medicine Subject: Re: PAULING, SZENT-GYORGYI, VITAMIN C AND ME Date: 8 May 1996 17:01:43 GMT Organization: UTHSCSA Lines: 154 Distribution: world Message-ID: <4mqk1n$hah@cosmos.uthscsa.edu> NNTP-Posting-Host: deneke.uthscsa.edu X-Newsreader: Xref: biosci bionet.general:21550 bionet.biology.cardiovascular:952 bionet.cellbiol:4639 bionet.glycosci:673 bionet.biophysics:1952 bionet.molbio.ageing:2702 bionet.molbio.proteins:7836 bionet.neuroscience:14004 sci.med:122778 sci.research.careers:10166 talk.politics.medicine:51285 In article , Bert Gold writes: >April 22, 1996 > >Walking back from the bank to the hospital today I was prompted to >buy several large juicy navel oranges. I've been eyeing oranges for many >weeks now, but my wife and I have a long standing disagreement about >who picks the best ones. I thought the oranges she chose at a fruit >stand, just outside Half Moon Bay yesterday, were puny; she again >suggested I always pick the oversized, dried up ones. >Today, though, my oranges were winners: Sweet, juicy and large; >just the way I like them. > >In the news the past several days, as you've probably heard, Vitamin C >requirements (RDAs, Required Daily Allowance, set by the National Research >Council, the nation's highest scientific authority) for adult males have >been revalued upward. Not modestly either: But three-fold, and still >the upper limit is poorly defined. It seems an intramural researcher >at NIH finally got the funding go ahead to do some of the experiments >which Linus Pauling and Albert Szent-Gyorgyi always longed to do >before the end of their lives, but were denied funding for. > >It is notable that in this latest work (1), seven men in their twenties >were fed increasing doses of Vitamin C after being starved for >it in their diet. Although they never showed signs of scurvy, >each volunteer did report feeling uncomfortable when deprived >of Vitamin C. The NIH authors note that because the subjects >of their study were young men in their 20's, the results are >limited to this subject group. > >The vivid image of Szent-Gyorgyi standing in front of Whitman Auditorium >in Woods Hole holding a bottle of glyoxyl in one hand and Vitamin C >in the other is permanently etched in my memory. I'll not easily >forget the hypothesis of 'radical scavenging' which Szent-Gyorgyi >put to us that day: He said that he and Pauling had been talking >about the central importance of OXYGEN in human metabolism >and explained that he had been concerned for a good deal of time about >reactions generating singlet oxygen. These, produced in abundance >during mitochondrial "charge transfer" in liver parenchyma, and other >aerobically active tissues, must absorb electrons pairwise in order to >avoid causing damage. > >In 1931, Pauling discovered the superoxide radical, which, though >produced in minute quantities as an unwanted byproduct of oxidative >phosphorylation, has enormous destructive capacity if not defused (2). >During lectures given at UC Berkeley in the 80s, Pauling paid >tribute to the remarkable specificity of the enzymatic system that >neutralizes it: Superoxide dismutase; at the same time, however, >he noted that this enzyme does not defuse the lesser, but still significant >intracellular destructive capacity of singlet oxygen or the hydroxide radical >(and unkown to him at the time, the NO radical). > >Superoxide dismutase transforms superoxide radicals to peroxide which can >then be eliminated as water after the action of peroxidase and catalase. >But what is the body to do with all the unpaired electrons such a process >would generate? Szent-Gyorgyi told that oxygen itself could absorb unpaired >electrons if only it were conjugated to carbon in the form of a double >carbonyl: the simplest such compound being glyoxyl, commonly found in human >liver, for this compound had sufficient electron delocalization to provide >something of an electron sink. And, the supply of glyoxyl is renewable >through the action of the glutathione-S-tranferase system, present in >liver mitochondria. Further, Szent-Gyorgyi told us excitedly, two >englishmen and a german, H.D. Dakin, H.W. Dudley and C. Neuberg had, in >1913 discovered that methylated-glyoxyl could be transformed to a potent >energy source itself, lactic acid, through the action of glyoxylase, >which they discovered in that year. So, in one fell swoop, Szent-Gyorgyi >had connected for us the relationship between oxygen radical and one >carbon metabolism. I promise I will discuss this relationship in the >next few weeks, when I write another essay, tentatively titled, >Folic Acid and I. > >In comments honoring Szent-Gyorgyi on the occasion of his 82nd >birthday, Linus Pauling recalled Vitamin C's discovery by Szent-Gyorgyi >in 1928. Pauling noted that Szent-Gyorgyi had written in 1939 (4) that >although organisms were generally well adapted to their surroundings, >that the destruction of the natural environment endangered that >adapatation. "I have a strong faith in the perfection of the human >body", Szent-Gyorgyi wrote in 1939, "and I think that vitamins are an >important factor in its coordination with its surroundings. Vitamins, >if properly understood and applied, will help us to reduce human suffering >to an extent which the most fantastic mind would fail to imagine." (3) > >On the day that I met him in Woods Hole, Szent-Gyorgyi told us he and Pauling >(three Nobel prizes) between them suggested that significant increases >in Vitamin C intake were almost certainly required by an adult body, >perhaps especially at times when oxygen production was increased. >Szent-Gyorgyi had insisted in a book he had written two years before, >that Vitamin C, especially when complexed with manganese in >the presence of oxygen, could synthesize a free radical form, as well and as >easily as it could form a dehydroascorbate, oxidized form in the >presence of copper or iron and in alliance with an enzyme he'd >discovered in plants in 1931 and named 'ascorbic acid oxidase.' >Szent-Gyorgyi insisted that it was in some sense the equilibrium between >the various oxidized and reduced forms of ascorbate which provided its >multifarious activities. > >And yet, this week, fully 18 years after the memorable talk by >Szent-Gyorgyi which I just described, the Vitamin C RDA for men >has been adjusted three-fold upward. > >It makes me wonder at the brazen inefficiency of the research >'establishment'... > >The knowledge that both Linus and Albe died >without ever attaining a modicum of funding for their final >nutrition project confirms some of my worst fears about >our ways of deciding what research is worthy in this >country. > >My hope in writing this is that our children will not suffer >because of the lack of wisdom of their leaders in >making decision about what to and what not to study. > >Bert Gold >San Francisco > >References > >(1) Levine, M.; Conry-Cantilena, C.; Wang, Y.; Welch, R.W.; Washko, P.W.; >Dhariwal, K.R.; Park, J.B.; Lazarev, A.; Graumlich, J.G., King, J.; >Cantilena, L.R. (1996) Vitamin C pharmacokinetics in healthy >volunteers: Evidence for a recommended dietary allowance. >Proc. Natl. Acad. Sci. USA; 93, 3704-3709. > >(2) Marinacci, B. (1995) Linus Pauling in His Own Words, New York, Simon & >Shuster. > >(3) Kaminer, B., ed. (1977) Search and Discovery, A tribute to Albert >Szent-Gyorgyi, New York, Academic Press. > >(4) Szent-Gyorgyi, A. (1939) On Oxidation, Fermentation, Vitamins, Health >and Disease. Baltimore, Williams and Wilkins. > > Pauling and Szent-Gyorgyi are my heros too, but neither a conspiracy or scientific incompetence is necessary to explain delays in jumping to recommend large doses of vitamin C. In fact chemistry, physiology etc. of ascorbic acid has been under intense investigation over the last several decades. It is well known that ascorbate, under some conditions acts a a pro-oxidant rather than an antioxidant. As for all nutritional additives it is extremely difficult to determine whether consumption of high levels of the pure product may cause toxic effects which are neutralized by other components present in the natural food sources. Do large doses of ascorbate protect, deplete, or block induction of other antioxidants? I can probably find instances of al