From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!bvc.edu!yuqian From: yuqian@bvc.edu (Suicidal Freshman) Newsgroups: bionet.molbio.proteins,bionet.molbio.rapd Subject: New Dedicated Bio-tech/science/chem BBS Message-ID: <1995May2.123420.10885@bvc.edu> Date: 2 May 95 12:34:20 CDT Organization: Buena Vista College, Storm Lake, IA Lines: 27 Xref: biosci bionet.molbio.proteins:4430 bionet.molbio.rapd:1118 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% A NEW BBS IS ONLINE AND NEEDS PEOPLE LIKE YOU TO HELP IT PROSPER The IDT Online Information System is now up and running and needs intelligent users such as yourselves. Anyone interested in Chemistry or Biology or any twist of the two is more than welcome. If you are a user of Oligonucleotides, there is an online order system that allows you to place your order directly into the production facility for 48 hour turnaround. THERE IS ABSOLUTELY NO CHARGE FOR USAGE. YOU DO NOT HAVE TO BE A CUSTOMER TO USE THE SYSTEM Just a sample of some of the discussion forums: Parsitology Classifieds Employment Chemical Engineering Computer Software Biochemistry Programming Molecular Medicine Genetics Organic Chemistry Oligo Design Scientific Humor BioPhysics Cell Biology Internet Topics Biomed Engineering Late Breaking News Immunology NeuroScience Suggestions The system is in place, and we are looking for good users, and a few Forum Moderators, so: Check it out: telnet 204.71.106.202 or telnet idtdna.com %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!howland.reston.ans.net!news.cac.psu.edu!news.pop.psu.edu!hudson.lm.com!netline-fddi.jpl.nasa.gov!nntp-server.caltech.edu!imppig!ranabir From: ranabir@imppig.caltech.edu (Ranabir Sinha Roy) Newsgroups: bionet.molbio.proteins Subject: Commercial source for PURE RNase-S PROTEIN Date: 3 May 1995 02:18:03 GMT Organization: California Institute of Technology, Pasadena Lines: 13 Message-ID: <3o6p4r$6ap@gap.cco.caltech.edu> NNTP-Posting-Host: imppig.caltech.edu Keywords: S-Protein, RNase X-Newsreader: NN version 6.5.0 #10 (NOV) Hi! I need pure S-protein (bovine pancreatic ribonuclease S-protein i.e. residues 20-124 of RNase-A) for crystallography experiments. The stuff from Sigma is at least 2 bands (1-2 kD apart) on an SDS phast-gel. Does anyone know of an alternate vendor for this protein? I am not interested in the ribonuclease-S COMPLEX. I just need the larger (S-protein) fragment...please e-mail me at sinharoy@romeo.caltech.edu with info/advice. Thanks! Ranabir Sinha Roy Grad. Student Dept. of Chemistry Caltech From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!nntp.uio.no!nac.no!Norway.EU.net!EU.net!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!uunet!newsflash.concordia.ca!nstn.ns.ca!ac.dal.ca!ac.dal.ca!nntp Newsgroups: bionet.molbio.proteins Subject: BCA Protein Assay and Detergents!! Message-ID: <1995May2.221027.37893@ac.dal.ca> From: JWChish@ac.dal.ca (Jeffrey W. Chisholm) Date: 2 May 95 22:10:26 -0300 Organization: Dalhousie University Nntp-Posting-Host: jchisholm.med.dal.ca X-Newsreader: WinVN 0.93.14 Lines: 41 Somebody recently posted that up to 1% SDS could be added to the BCA protein system for the measurement of lipid rich protein samples. As I work on lipoproteins and was in the process of switching to the BCA system from one of the modified Lowry methods (With SDS or Deoxycholate), this was of interest to me. However, in my hands, 1% SDS in the BCA assay yielded significantly lower protein values in VLDL, LDL and other lipid rich particles as compared with the plain BCA protocol. In addition, when using the micro BCA method (Pierce) with microtiter plates, the SDS was prone to precipitation with samples containing high salt. As a result, I tried the BCA assay adding 1%, .5% and 0.25% Deoxycholate as a solubilizing agent. Overall, 0.25% appears to work best, yielding a std. curve (BSA) identical to the plain BCA protocol and giving significantly larger protein mass values in lipid rich particles and comparable values in lipid depleted samples. 1% and 0.5% DOC yielded protein masses equal to or less than the plain BCA kit. From now on, I will be using the BCA protocol with 0.25% DOC for regular determinations of protein in lipid rich samples. Incidently, 0.25% DOC was the concentration used in our DOC modified Lowry procedure. For those not familiar with the BCA protocol, it has a working range from 10- 2000 ug/ml and yields a slightly non-linear curve, such that I usually construct two lines for interpolating samples, eg 10-200 ug/ml std. set and 200-2000 ug/ml std. set. For those wishing only to construct one curve, the non-linear nature of the curve seems to be reduced upon extended incubation of the reaction (beyond 30 min at 37 Celsius). Hope some of you find this useful, Comments welcome, Jeffrey W. Chisholm Lipoprotein Research Group Dept. of Biochemistry Dalhousie University Halifax, N.S., Can. Email: JWChish@ac.dal.ca From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!bvc.edu!yuqian From: yuqian@bvc.edu (Suicidal Freshman) Newsgroups: bionet.molbio.proteins,bionet.molbio.rapd Subject: New Dedicated Bio-tech/science/chem BBS Message-ID: <1995May2.123306.10882@bvc.edu> Date: 2 May 95 12:33:06 CDT Organization: Buena Vista College, Storm Lake, IA Lines: 27 Xref: biosci bionet.molbio.proteins:4429 bionet.molbio.rapd:1117 %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% A NEW BBS IS ONLINE AND NEEDS PEOPLE LIKE YOU TO HELP IT PROSPER The IDT Online Information System is now up and running and needs intelligent users such as yourselves. Anyone interested in Chemistry or Biology or any twist of the two is more than welcome. If you are a user of Oligonucleotides, there is an online order system that allows you to place your order directly into the production facility for 48 hour turnaround. THERE IS ABSOLUTELY NO CHARGE FOR USAGE. YOU DO NOT HAVE TO BE A CUSTOMER TO USE THE SYSTEM Just a sample of some of the discussion forums: Parsitology Classifieds Employment Chemical Engineering Computer Software Biochemistry Programming Molecular Medicine Genetics Organic Chemistry Oligo Design Scientific Humor BioPhysics Cell Biology Internet Topics Biomed Engineering Late Breaking News Immunology NeuroScience Suggestions The system is in place, and we are looking for good users, and a few Forum Moderators, so: Check it out: telnet 204.71.106.202 or telnet idtdna.com %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!ns1.faseb.org!darwin.sura.net!nih-csl!loglady.ninds.nih.gov!johnk From: johnk@spasm.niddk.nih.gov (John Kuszewski) Subject: Well, that about wraps it up for God. Message-ID: <1995May2.163638.26210@alw.nih.gov> Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: spasm.niddk.nih.gov Reply-To: johnk@spasm.niddk.nih.gov (John Kuszewski) Organization: National Insts. of Health Date: Tue, 2 May 1995 16:36:38 GMT Lines: 23 In case you haven't heard, George Rose has solved the folding problem :-/ (He wouldn't say so yet, but he has.) Anyone know of any other good lines of work? -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || John Kuszewski || |/ /| || johnk@spasm.niddk.nih.gov || / /|| || \/ / / || \/ that's MISTER protein G to you! |/__/| | /_________| "Biophysics has driven me to an attitude of apocalyptic doom" --Frank Delaglio From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!ux1.cso.uiuc.edu!elarson From: elarson@ux1.cso.uiuc.edu (larson eric) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: galactose/galactose oxidase assay information needed Date: 3 May 1995 04:14:10 GMT Organization: University of Illinois at Urbana Lines: 39 Message-ID: <3o6vui$t6g@vixen.cso.uiuc.edu> References: NNTP-Posting-Host: ux1.cso.uiuc.edu Xref: biosci bionet.molbio.methds-reagnts:27867 bionet.molbio.proteins:4435 bipin dalmia writes: >hi: i need to set-up an assay for galactose using the galactose oxidase, >peroxidase method. would need any references to such an assay. also, does >anyone know if a commercial kit is available with preweighed enzymes etc. >similar to the one from Sigma for glucose assay? Sigma does not have a >galactose kit. if you can send any information, i would appreciate very >much. either post, or e-mail to dalmiabk@phibred.com. thanx. ============= The Worthington enzyme catalog lists a procedure for assaying Galactose Oxidase. The book lists a review (Malmstron et al. (1975) The Enzymes, XIIB, 3rd. Ed. (Boyer, P. D., ed.) pg 527). I suggest starting here (either the Worthington catalog or The Enzymes). The procedure is fairly easy, but I don't know if anyone still produces the kits (the Worthington catalog states they do, but it is an old catalog -- if you've ever seen a Wothington catalog you'd understand why people save these as reference works). I've done the assay -- it's relatively straightforward (typical assay with lots of additions). I do suggest caution if you're going to have glycerol around. There is a very low activity with glcyerol (makes glyceraldehyde) that can futz up long-term incubations (easy to assay this activity -- just have 20% glycerol in your assay and you should get substantial activity without galactose being present. Ship me a fax number and I'll send you the two pages of the Worthington catalog (and an ancient page of typed notes -- which I have no intention of typing in this evening). -- Eric Larson | University of Illinois at Urbana-Champaign USDA/Agronomy | 190 PABL; 1201 W. Gregory; Urbana, IL 61801 elarson@ux1.cso.uiuc.edu | Voice 217.244.3079 Fax 217.244.4419 Fidonet: 1:233/4.1 | My opinions are my own, but correct :-) From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Path: biosci!daresbury!trane.uninett.no!nac.no!Norway.EU.net!EU.net!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!gw1.phibred.com!dalmiabkmac.phibred.com!dalmiabk From: bipin dalmia Subject: galactose/galactose oxidase assay information needed Message-ID: X-Xxmessage-Id: X-Xxdate: Tue, 2 May 95 16:09:10 GMT Sender: news@phibred.com (USENET News System) Organization: pioneer hi-bred X-Useragent: Version 1.1.3 Date: Tue, 2 May 1995 21:15:59 GMT Lines: 8 Xref: biosci bionet.molbio.methds-reagnts:27856 bionet.molbio.proteins:4432 hi: i need to set-up an assay for galactose using the galactose oxidase, peroxidase method. would need any references to such an assay. also, does anyone know if a commercial kit is available with preweighed enzymes etc. similar to the one from Sigma for glucose assay? Sigma does not have a galactose kit. if you can send any information, i would appreciate very much. either post, or e-mail to dalmiabk@phibred.com. thanx. bip From owner-proteins@net.bio.net Mon May 01 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!nntp.uio.no!nac.no!Norway.EU.net!EU.net!howland.reston.ans.net!gatech!willis.cis.uab.edu!maze.dpo.uab.edu!news!zm00025 From: zm00025@uabdpo.dpo.uab.edu (Jon Grim) Newsgroups: bionet.molbio.proteins Subject: Help on KDEL signal needed Followup-To: bionet.molbio.proteins Date: Tue, 2 May 95 17:44:52 GMT Organization: University of Alabama at Birmingham Lines: 12 Message-ID: NNTP-Posting-Host: 138.26.196.125 X-Newsreader: VersaTerm Link v1.1 I am adding a KDEL to the end of a protein that I also want to detect with an antibody to a TAG fragment. Will the order of these be of importance? Or, will the KDEL work if I put it before (5') of the TAG? Or does a KDEL have to be the very last part of a protein to work?? Anyhelp is appreciated, Please email personally if possible. Jon Grim University of Alabama-Birmingham From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!pipex!lyra.csx.cam.ac.uk!moose.bioc.cam.ac.uk!user From: nef@mole.bio.cam.ac.uk (Nick Fisher) Newsgroups: bionet.molbio.proteins Subject: Re: Well, that about wraps it up for God. Date: Wed, 03 May 1995 12:42:11 +0000 Organization: Cambridge University (Biochemistry Department) Lines: 9 Message-ID: References: <1995May2.163638.26210@alw.nih.gov> NNTP-Posting-Host: moose.bioc.cam.ac.uk In article <1995May2.163638.26210@alw.nih.gov>, johnk@spasm.niddk.nih.gov (John Kuszewski) wrote: > In case you haven't heard, George Rose has solved the > folding problem :-/ > > (He wouldn't say so yet, but he has.) hmm...it doesn't involve palladium electrodes and heavy water does it? ;) From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!swrinde!pipex!sunic!sunic.sunet.se!news.uni-c.dk!eagle.novo.dk!usenet From: Lauge Schaffer Newsgroups: bionet.molbio.proteins Subject: Re: CNBr -Sepharose 4B Date: 3 May 1995 14:13:54 GMT Organization: Novo Nordisk Lines: 30 Message-ID: <3o8332$c9l@eagle.novo.dk> References: <3nogoq$p25@mercury.hgmp.mrc.ac.uk> NNTP-Posting-Host: lsc.novo.dk Why should urea or Gu interfere with your immobilization? Neither contains "real" amino groups, i.e. amino groups which would block CNBr activated Sepharose. Lauge Schaffer (anonymous) wrote: > > I am looking for some help in coupling a poorly soluble protein to CNBr- > activated Sepharose 4B. We want to affinity purify some antibodies, but > the antigen (a recombinant protein) is only soluble in guanidine HCl or > urea. When diluted at least 20 fold with phosphate, the protein remains in > solution, but at these concentrations the coupling efficiency will be very low. > > I was wondering if anyone has any experience of coupling a protein as a > slurry, perhaps in the presence of a non-ionic detergent. Alternatively, > is there an affinity matrix which is compatable with solvents containing > amino groups. > > > Thanks in advance > > Pete Jenner > > Dept of Mycobacterial Res. > NIMR. > London > UK. From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!rutgers!gatech!howland.reston.ans.net!Germany.EU.net!EU.net!uknet!bcc.ac.uk!jones From: jones@bsm.bioc.ucl.ac.uk (David Jones) Subject: Re: Well, that about wraps it up for God. Message-ID: <1995May3.150119.24671@ucl.ac.uk> Date: Wed, 3 May 1995 15:01:19 GMT References: <1995May2.163638.26210@alw.nih.gov> Organization: University College London X-Newsreader: TIN [version 1.2 PL2] Lines: 22 John Kuszewski (johnk@spasm.niddk.nih.gov) wrote: > In case you haven't heard, George Rose has solved the > folding problem :-/ > (He wouldn't say so yet, but he has.) > Anyone know of any other good lines of work? I don't blame George Rose for holding back such a claim. To keep on the Douglas Adams track, it would be very impressive indeed for someone to solve the protein folding problem today - not because they would have found the answer (42?) but because it would imply that they know what the question is! >---------------------------------------------------------------------------< This message was written, produced and executively directed by Dr David Jones Address: Department of Biochemistry and | Email: jones@bsm.bioc.ucl.ac.uk Molecular Biology, University College, | Tel: +44 171 387 7050 x3879 Gower Street, London WC1E 6BT, U.K. | Fax: +44 171 380 7193 Disclaimer: STANDARD > KEYWORDS : OPINIONS MY OWN NOBODY ELSE'S WHATSOEVER From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!rutgers!gatech!swrinde!pipex!lyra.csx.cam.ac.uk!mole.bio.cam.ac.uk!smb18 From: smb18@mole.bio.cam.ac.uk (Simon Brocklehurst (Bioc)) Newsgroups: bionet.molbio.proteins Subject: Re: Well, that about wraps it up for God. Date: 3 May 1995 13:02:11 GMT Organization: University of Cambridge, England Lines: 21 Message-ID: <3o7usj$21@lyra.csx.cam.ac.uk> References: <1995May2.163638.26210@alw.nih.gov> NNTP-Posting-Host: mole.bio.cam.ac.uk johnk@spasm.niddk.nih.gov (John Kuszewski) writes: >In case you haven't heard, George Rose has solved the >folding problem :-/ >(He wouldn't say so yet, ______________________ ... Why, then, are _you_ suggesting that he has? > but he has.) ____________ ... Does George Rose know you are making this claim on his behalf? _________________________________________________________________________ | | ,_ o Simon M. Brocklehurst, | / //\, Oxford Centre for Molecular Sciences | \>> | Department of Biochemistry, University of Oxford, | \\, Oxford, UK. | E-mail: smb@bioch.ox.ac.uk |________________________________________________________________________ From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!daresbury!trane.uninett.no!sunic!sunic.sunet.se!seunet!news2.swip.net!doc.news.pipex.net!pipex!uknet!bcc.ac.uk!jones From: jones@bsm.bioc.ucl.ac.uk (David Jones) Subject: Re: Comment to S.Blocklehurst (Re: Well, ...) Message-ID: <1995May3.224640.20424@ucl.ac.uk> Date: Wed, 3 May 1995 22:46:40 GMT References: <950503170409.2040beb4@scri.fsu.edu> Organization: University College London X-Newsreader: TIN [version 1.2 PL2] Lines: 42 VICTOR B. STRELETS (STRELETS@SCRI.FSU.EDU) wrote: > Simon, > I think that John was correct making such comments about work > of G.Rose. This was not a claim that folding is really solved, > but rather comment about his personal impression from Rose's work. > Imagine: already half a year we hear something about this > intriguing work, but any description of the algorithm is still > absent.. Rose's result is principal for the large area of biocomputing > problems, and too many people are waiting for a paper or > at least for personal comments from lucky visitors of the > rare Rose's presentations. > As far I know, John commented his impression immediately after > presentation of G.Rose results on some sort of seminar at NIH. > What is wrong with that? And for me this is interesting because > I would like to know (in absense of any possibility to visit > such a seminar in other part of US) how different people become > convinced in the efficacy of this algorithm.. Of course, this is > only while I'm waiting for publication/description. > And John's comment shows something, isn't it? > STR Unfortunately there seem to be three diverse groups of people here. The first group seem to have been to one of George Rose's seminars. The second group have heard some rumours and have not been to one of George Rose's seminars - but would like to. The third group (which certainly includes me) have heard absolutely nothing about this supposed breakthrough - until now. Perhaps I'm the only one in the third group? Hands up all those who have heard about this work by George Rose... oh dear, how embarassing - just me, then! Just to make some use of the wasted bandwidth here - why doesn't someone in the first group let us in the third group in on what's supposed to be going on. If he's spoken about this at a seminar then a brief synopsis isn't going to hurt anyone - then we can _all_ enjoy the suspense until the paper comes out! >---------------------------------------------------------------------------< This message was written, produced and executively directed by Dr David Jones Address: Department of Biochemistry and | Email: jones@bsm.bioc.ucl.ac.uk Molecular Biology, University College, | Tel: +44 171 387 7050 x3879 Gower Street, London WC1E 6BT, U.K. | Fax: +44 171 380 7193 Disclaimer: STANDARD > KEYWORDS : OPINIONS MY OWN NOBODY ELSE'S WHATSOEVER From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!umn.edu!limerick.cbs.umn.edu!hiroki From: hiroki@limerick.cbs.umn.edu (Hiroki Morizono) Subject: Re: Well, that about wraps it up for God. Message-ID: Sender: news@news.cis.umn.edu (Usenet News Administration) Nntp-Posting-Host: limerick.cbs.umn.edu Reply-To: hiroki@limerick.cbs.umn.edu Organization: University of Minnesota, Twin Cities X-Newsreader: TIN [version 1.2 PL2] References: <1995May2.163638.26210@alw.nih.gov> Date: Thu, 4 May 1995 00:08:07 GMT Lines: 8 John Kuszewski (johnk@spasm.niddk.nih.gov) wrote: : In case you haven't heard, George Rose has solved the : folding problem :-/ : (He wouldn't say so yet, but he has.) : Anyone know of any other good lines of work? How about hunting for the Great Pumpkin? Hiroki From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!news.iastate.edu!baker From: baker@iastate.edu (Wayne R. Baker) Newsgroups: bionet.molbio.proteins Subject: A Plea To John K! (was Re: Comment to S.Blocklehurst) Date: 4 May 1995 02:49:02 GMT Organization: Biochemistry & Biophysics, Iowa State University Lines: 34 Message-ID: <3o9fau$9a3@news.iastate.edu> References: <950503170409.2040beb4@scri.fsu.edu> <1995May3.224640.20424@ucl.ac.uk> NNTP-Posting-Host: pv0a05.vincent.iastate.edu In bionet.molbio.proteins, David Jones wrote : :Unfortunately there seem to be three diverse groups of people here. :The first group seem to have been to one of George Rose's seminars. :The second group have heard some rumours and have not been to one of :George Rose's seminars - but would like to. :The third group (which certainly includes me) have heard absolutely :nothing about this supposed breakthrough - until now. Perhaps I'm the :only one in the third group? Hands up all those who have heard about :this work by George Rose... oh dear, how embarassing - just me, then! You're not the only one, although I'm a wee bit behind in my journals. :-( :Just to make some use of the wasted bandwidth here - why doesn't :someone in the first group let us in the third group in on what's :supposed to be going on. If he's spoken about this at a seminar then :a brief synopsis isn't going to hurt anyone - then we can _all_ :enjoy the suspense until the paper comes out! I second the motion. All in favor...? Opposed...? The ayes have it! Rose did publish something on helix forming propensities in "Proteins" last year, but the work is now nearly two years old. Last March, Scheraga seemed to think he was close to a solution. Levitt also seemed like he was on to something as well. At the Beckman conference last June, Patrick Argos showed some results for simple systems that were pretty impressive. I'd be interested in how Rose's algorithm compares. Anyone care to scoop Dr. Rose? :-) Wayne Baker (baker@iastate.edu) Maybe a great magnet pulls Biochemistry & Biophysics All souls towards truth Iowa State University -- k. d. lang, "Constant Craving" From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!news.iastate.edu!baker From: baker@iastate.edu (Wayne R. Baker) Newsgroups: bionet.molbio.proteins Subject: Re: Predicting pKas from PDBs Date: 4 May 1995 02:09:38 GMT Organization: Biochemistry & Biophysics, Iowa State University Lines: 25 Message-ID: <3o9d12$8dk@news.iastate.edu> References: <1995May3.183936.21837@alw.nih.gov> NNTP-Posting-Host: pv0a05.vincent.iastate.edu In bionet.molbio.proteins, John Kuszewski wrote :Does anyone know if there's a method to predict :the pKas of ionizable protein groups from :atomic coordinates? Quite a few, actually, although the accuracy is another matter. I believe DelPhi from Honig's group can do the calculations, which is available for a small fee via anon ftp and also is in the package Insight. Bashford et al [(1993) Biochemistry vol 32 p8055] has a program called MEAD that is available for free via anon ftp. The reference has the info on how to get it. I'm hoping to install it Real Soon so I can see how well it does. Those are the only packages I know of that are available for outside users. Warshel has a program called POLARIS [Warshel et al (1984) PNAS vol 81 p4785] and there is an FDPB method described by Antosiewicz [(1994) vol 238 p415] that did a fairly good job on RNase A. And the ubiquitous Fred Richards did this ages ago [Matthews & Richards (1982) Biochemistry vol. 21 p4989]. I don't know if any of these folks have made their packages available for academic use; if you find out, lemme know. :-) Wayne Baker (baker@iastate.edu) Maybe a great magnet pulls Biochemistry & Biophysics All souls towards truth Iowa State University -- k. d. lang, "Constant Craving" From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!biosci!not-for-mail From: xhu1@ux1.cso.uiuc.edu (hu xiche) Newsgroups: bionet.software.gcg,bionet.molbio.proteins,bionet.molec-model Subject: Z-Score and cutoff in protein sequence alignment Date: 3 May 1995 18:46:15 -0700 Organization: University of Illinois at Urbana Lines: 13 Sender: biohelp@net.bio.net Distribution: world Message-ID: <3o8j17$jrs@vixen.cso.uiuc.edu> NNTP-Posting-Host: net.bio.net Summary: Z-Score, Protein Keywords: sequence alignment Xref: biosci bionet.software.gcg:1162 bionet.molbio.proteins:4446 bionet.molec-model:351 I am using the BESTFIT program in the GCG package to align two protein sequences. I would like to evaluate the statistical significance of such an alignment. By using the -RAND option, I can get the averaged quality of the "shuffled" random sequences and the standard deviation from the output of BESTFIT. But I don't know how to convert them into "Z-score". I will deeply appreciate your help in answering the following questions: (1) How should the "Z-Score" be calculated? and (2) What is the cutoff for statistically meaningful alignment in terms of "Z-Score"? Any references to published journal articles or names of experts who will be able to help will be very helpful. Thanks in advance, Xiche Hu From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!uunet!zib-berlin.de!fu-berlin.de!novell-compaq.rz-berlin.mpg.DE!not-for-mail From: Schnibbe@MPIMG-Berlin-Dahlem.MPG.DE (Thomas Schnibbe) Newsgroups: bionet.molbio.proteins,bionet.molec-model Subject: Conformation of peptides in aqueous solution? Date: Wed, 3 May 1995 19:39:36 GMT +0100 Organization: Max Planck Institut fuer molekulare Genetik, Berlin Lines: 28 Message-ID: NNTP-Posting-Host: novell-compaq.rz-berlin.mpg.de (141.14.129.150) X-Access: 16 25 816 X-Newsreader: Trumpet for Windows [Version 1.0 Rev B] Xref: biosci bionet.molbio.proteins:4445 bionet.molec-model:350 Dear Netters, I have a question concerning peptide conformation in aqueous solution. It has been shown that secondary structure formation in aqueous solution should be negligible for all short peptides (shorter than 13 - 15 aa or so). Increased chain length moves along with secondary structure formation: Medium size peptides may present a population with secondary structure in a rapid equilibrium with random, mainly extended, conformations. Large peptides/proteins beyond a certain chain length show stable (super-) secondary and tertiary structures (quite stable and somehow dynamic, too). My question is: WERE (AT WHICH SIZE) IS THE APPROXIMATE TRANSITION "DYNAMIC - STABLE"? (aqueous solution no detergents etc., room-temperature) (Of course, secondary/tertiary structure formation strongly depends on physico-chemical properties of the bulk solution, the primary structure, temperature etc., too) Since I'm working on synthetic peptides you would help me a lot. Thank you for your time, Thomas From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!gatech!bloom-beacon.mit.edu!mojo.eng.umd.edu!cs.umd.edu!news.umbc.edu!haven.umd.edu!purdue!mozo.cc.purdue.edu!fspc62.foodsci.purdue.edu!MURIANAP From: MURIANAP@FOODSCI.PURDUE.EDU (Pete Muriana) Newsgroups: bionet.molbio.proteins Subject: Commercial supplier of antibodies against Salmonella fimbriae? Date: Wed, 3 May 1995 08:20:40 LOCAL Organization: FOOD SCIENCE DEPT./PURDUE UNIVERSITY Lines: 11 Message-ID: NNTP-Posting-Host: fspc62.foodsci.purdue.edu X-Newsreader: Trumpet for Windows [Version 1.0 Rev B final beta #4] Does anyone know of a commercial supplier of antibodies to Salmonella fimbrial antigens? -Regards, PEter ********************************************************************** * Peter M. Muriana, Ph.D. 317-494-8284 TEL * * Dept. of Food Science 317-494-7953 FAX * * Purdue University murianap@foodsci.purdue.edu * * Smith Hall * * W. Lafayette, IN 47907-1160 * ********************************************************************** From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!SCRI.FSU.EDU!STRELETS From: STRELETS@SCRI.FSU.EDU ("VICTOR B. STRELETS") Newsgroups: bionet.molbio.proteins Subject: Comment to S.Blocklehurst (Re: Well, ...) Date: 3 May 1995 14:04:24 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 19 Sender: daemon@net.bio.net Distribution: world Message-ID: <950503170409.2040beb4@scri.fsu.edu> NNTP-Posting-Host: net.bio.net Simon, I think that John was correct making such comments about work of G.Rose. This was not a claim that folding is really solved, but rather comment about his personal impression from Rose's work. Imagine: already half a year we hear something about this intriguing work, but any description of the algorithm is still absent.. Rose's result is principal for the large area of biocomputing problems, and too many people are waiting for a paper or at least for personal comments from lucky visitors of the rare Rose's presentations. As far I know, John commented his impression immediately after presentation of G.Rose results on some sort of seminar at NIH. What is wrong with that? And for me this is interesting because I would like to know (in absense of any possibility to visit such a seminar in other part of US) how different people become convinced in the efficacy of this algorithm.. Of course, this is only while I'm waiting for publication/description. And John's comment shows something, isn't it? STR From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!rutgers!gatech!news-feed-1.peachnet.edu!darwin.sura.net!nih-csl!loglady.ninds.nih.gov!johnk From: johnk@spasm.niddk.nih.gov (John Kuszewski) Subject: Predicting pKas from PDBs Message-ID: <1995May3.183936.21837@alw.nih.gov> Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: spasm.niddk.nih.gov Reply-To: johnk@spasm.niddk.nih.gov (John Kuszewski) Organization: National Insts. of Health Date: Wed, 3 May 1995 18:39:36 GMT Lines: 20 Does anyone know if there's a method to predict the pKas of ionizable protein groups from atomic coordinates? -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || John Kuszewski || |/ /| || johnk@spasm.niddk.nih.gov || / /|| || \/ / / || \/ that's MISTER protein G to you! |/__/| | /_________| "Biophysics has driven me to an attitude of apocalyptic doom" --Frank Delaglio From owner-proteins@net.bio.net Tue May 02 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bloom-beacon.mit.edu!gatech!news-feed-1.peachnet.edu!darwin.sura.net!nih-csl!loglady.ninds.nih.gov!johnk From: johnk@spasm.niddk.nih.gov (John Kuszewski) Subject: Re: Well, that about wraps it up for God. Message-ID: <1995May3.151539.18056@alw.nih.gov> Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: spasm.niddk.nih.gov Reply-To: johnk@spasm.niddk.nih.gov (John Kuszewski) Organization: National Insts. of Health References: <1995May2.163638.26210@alw.nih.gov> <3o7usj$21@lyra.csx.cam.ac.uk> Date: Wed, 3 May 1995 15:15:39 GMT Lines: 38 In article <3o7usj$21@lyra.csx.cam.ac.uk>, smb18@mole.bio.cam.ac.uk (Simon Brocklehurst (Bioc)) writes: |> johnk@spasm.niddk.nih.gov (John Kuszewski) writes: |> |> >In case you haven't heard, George Rose has solved the |> >folding problem :-/ |> |> >(He wouldn't say so yet, |> ______________________ ... Why, then, are _you_ suggesting that he has? |> |> > but he has.) |> ____________ ... Does George Rose know you are making this claim on his |> behalf? I was an idiot for posting that yesterday. Obvisouly, I don't speak for George Rose, and I'm sorry if people thought I did. I do think that he has some extremely clever new stuff, though I'll wait for the paper before commenting again. -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || John Kuszewski || |/ /| || johnk@spasm.niddk.nih.gov || / /|| || \/ / / || \/ that's MISTER protein G to you! |/__/| | /_________| "Biophysics has driven me to an attitude of apocalyptic doom" --Frank Delaglio From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!rutgers!gatech!usenet.eel.ufl.edu!noc.netcom.net!news.sprintlink.net!psgrain!charnel.ecst.csuchico.edu!csusac!csus.edu!news.ucdavis.edu!library.ucla.edu!info.ucla.edu!nnrp.info.ucla.edu!usenet From: arne@hodgkin.mbi.ucla.edu (Arne Elofsson) Newsgroups: bionet.molbio.proteins Subject: Re: NO. Re: A Plea To John K! (was Re: Comment to S.Blocklehurst) Date: 04 May 1995 15:33:29 -0700 Organization: University of California, Los Angeles Lines: 31 Message-ID: References: <950503170409.2040beb4@scri.fsu.edu> <3o9fau$9a3@news.iastate.edu> <1995May4.152111.11523@alw.nih.gov> <1995May4.171649.29042@ucl.ac.uk> <3ob7h5$fjn@nexus.uiowa.edu> NNTP-Posting-Host: hodgkin.mbi.ucla.edu In-reply-to: kpmurphy@blue.weeg.uiowa.edu's message of 4 May 1995 18:48:05 GMT X-Newsreader: (ding) Gnus v0.63 In article <3ob7h5$fjn@nexus.uiowa.edu> kpmurphy@blue.weeg.uiowa.edu (murphy kenneth p) writes: > I agree with John that details should wait until the paper is published, which > will be June or July in Proteins. I will say that the approach they're using > is a build-up procedure using Monte Carlo and a simplified potential function. > I was very impressed with what Rose showed and personally feel that John's > estimate of ten years is too conservative. > ahh... Sounds kinda interesting. I saw some people doing this 3 years ago with a very complicated energy function, (some kind of ab initio) Of course it did not work. Anyone remember who that can have been ? I have a vague memore that they were european (german ?) So what else (besides the build up procedure) is new in Rose's stuff. compared to their 1994 work ? Same pot-fkn ? I guess they tried it on something bigger than a helix ? arne -- -------------------------------------------------------- From: Arne Elofsson Email: arne@hodgkin.mbi.ucla.edu WWW: http://www.doe-mbi.ucla.edu/arne/main.html From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!rutgers!gatech!swrinde!howland.reston.ans.net!news.cac.psu.edu!news.tc.cornell.edu!newsserver.sdsc.edu!nic-nac.CSU.net!charnel.ecst.csuchico.edu!csusac!csus.edu!news.ucdavis.edu!library.ucla.edu!info.ucla.edu!nnrp.info.ucla.edu!usenet From: arne@hodgkin.mbi.ucla.edu (Arne Elofsson) Newsgroups: bionet.molbio.proteins Subject: Re: Comment to S.Blocklehurst (Re: Well, ...) Date: 04 May 1995 15:24:35 -0700 Organization: University of California, Los Angeles Lines: 34 Message-ID: References: <950503170409.2040beb4@scri.fsu.edu> <1995May3.224640.20424@ucl.ac.uk> NNTP-Posting-Host: hodgkin.mbi.ucla.edu In-reply-to: jones@bsm.bioc.ucl.ac.uk's message of Wed, 3 May 1995 22:46:40 GMT X-Newsreader: (ding) Gnus v0.63 In article <1995May3.224640.20424@ucl.ac.uk> jones@bsm.bioc.ucl.ac.uk (David Jones) writes: Well spoken David. I have never heard about D.Rose's work either. And anything that is given to me in second hand is just to disregard. There are so many ways to cheat in this business, or atleast overinterpretting the data. To be quite frank even most first hand impressions from seminars can be very missleading. You have really to read the papers to see what people are doing. Every meeting I go to there is one or a couple posters that claims that they have solved the protein folding problem. Usually are these posters filled with some theoretical-computer-science-ai-crap. And then one table of data to show how close they can predict structures. Sometimes there is a small email adress at the bottom trying to sell you a program also. Well I guess we are not enough people out there that know that most of this is crap. arne PS. I know what thew question is, so if you know the answer 42, we should be happy. My question is to given any sequence build a model to within 1.0 A from a crustal structure. (At the moment I would be happu with 1.5 or even 2. (-: ) -- -------------------------------------------------------- From: Arne Elofsson Email: arne@hodgkin.mbi.ucla.edu WWW: http://www.doe-mbi.ucla.edu/arne/main.html From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!unb.ca!savidge From: savidge@unb.ca ("Dr. R. A. Savidge") Newsgroups: bionet.molbio.proteins Subject: glycoproteins Date: 4 May 1995 08:03:45 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <199505041203034207.savidge@unb.ca> Reply-To: savidge@unb.ca NNTP-Posting-Host: net.bio.net Can anyone tell me if there is an accurate way to estimate the molecular weight of a glycoprotein by SDS-PAGE? For example, are there glycoprotein marker kits available? ********************************************************************** Rod Savidge, PhD | E-mail: savidge@unb.ca Faculty of Forestry and \|/ Environmental Management \ | / Phone: (506) 453-4919 University of New Brunswick _\/ | \/_ Fredericton, NB CANADA \|/ Fax: (506) 453-3538 E3B 6C2 | From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!unb.ca!savidge From: savidge@unb.ca ("Dr. R. A. Savidge") Newsgroups: bionet.molbio.proteins Subject: glycoproteins Date: 4 May 1995 07:49:51 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1031 Sender: daemon@net.bio.net Distribution: world Message-ID: <199505041148316932.savidge@unb.ca> Reply-To: savidge@unb.ca NNTP-Posting-Host: net.bio.net --==_19950504114831_9387 Content-Type: text/plain; charset="us-ascii" Can anyone tell me if there is an accurate way to estimate the molecular weight of a glycoprotein by SDS-PAGE? For example, are there glycoprotein marker kits available? ********************************************************************** Rod Savidge, PhD | E-mail: savidge@unb.ca Faculty of Forestry and \|/ Environmental Management \ | / Phone: (506) 453-4919 University of New Brunswick _\/ | \/_ Fredericton, NB CANADA \|/ Fax: (506) 453-3538 E3B 6C2 | --==_19950504114831_9387 Content-Type: application/octet-stream; charset="us-ascii"; name="OXYGENFI." 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H!!!!!!!?!ehz!!L6!!!!!!!\!!@!!!!!!!!!!!!! H!!! PICT! !!!!!h!s!!!!!!!\!4!\!!!]! userdict /_cv known not currentdict /bu known and{bu}{save}ifelse userdict /_cv known not dup {userdict /_cv 20 dict put}if _cv begin /bdf{bind def}bind def /setcmykcolor where{/setcmykcolor get /cvcmyk exch def}{/cvcmyk{1 sub 4 1 roll 3{3 index add neg dup 0 lt{pop 0}if 3 1 roll}repeat setrgbcolor pop}bdf }ifelse /stg{cf ca /cs load setscreen setgray}bdf /strgb{cf ca /cs load setscreen setrgbcolor}bdf /stcmyk{cf ca /cs load setscreen cvcmyk}bdf /min1{dup 0 eq{pop 1}if}bdf currentscreen/cs exch def/ca exch def/cf exch def end currentdict /bn known and{bn}{restore}ifelse !!!!!!!!d!0drw2!!!e!!!l!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! !!!!!!!!!Q!f!!!m!!!!!d!`drw2!!!d!!!s!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!d!!!s!!!!!!!!!!!!!!!!!!!!!!!Z!!!!!!!!!!!!!!!!!!!!!!! !!!!!!!!!"!d!!!!!!!d!1drw2!!!!!T!!!^!X*!!X*!!!!!!!!!!!!!!!!!!!!!!! !!!.!!!!!!!(!!!U!5!!!!!!!d! drw2!:!!! !!!!!!!!!!!d!!drw2!:!!!!!!!!!!!!!!!!!d!1drw2!!!!!5!!!?!X*!!X*!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!d!!drw2!:!!!!!!!!!!!!!d!!drw2!:!!!$!!!!!!!d!4drw2!:!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! --==_19950504114831_9387-- From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!rutgers!gatech!howland.reston.ans.net!news.sprintlink.net!malgudi.oar.net!hyperion.wright.edu!desire.wright.edu!s012msg From: s012msg@desire.wright.edu (MARC S. GREENBERG) Newsgroups: bionet.molbio.proteins Subject: plasma protein binding Ques. Message-ID: <1995May4.173624.38742@desire.wright.edu> Date: 4 May 95 17:36:24 EST Organization: Wright State University Lines: 11 I need to develop or use a method to measure the binding of chlorinated acids (e.g., trichloroacetic acid = TCA) to plasma proteins. I have done extensive literature searches and it seems that the two most common methods for such measurements are equilibrium dialysis and ultra-filtration. I would like some suggestions as to other methods that may be useful to these protein-binding determinations. Are there any electrophoretic techniques that may be useful? Thanks in advance, Marc From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!PORTIA.CALTECH.EDU!EARN From: EARN@PORTIA.CALTECH.EDU (JUST ERIC) Newsgroups: bionet.molbio.proteins Subject: Yeast protease inhibitors Date: 4 May 1995 14:59:34 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HQ3XX4F53SBLD26P@PORTIA.CALTECH.EDU> NNTP-Posting-Host: net.bio.net I'm looking for help in creating an effective protease inhibitor cocktail for yeast extract - I am expressing a small soluble enzyme in Saccharomyces with a C-terminal affinity tag. I get plenty of expression, but at each step in the purification I see significant amounts of active enzyme which has lost the tag, as confirmed by SDS-PAGE. The inhibitors I am using in mybreaking buffer (for BeadBeater disruption) are: 5 mM phenanthroline 1mM pefablock 10 microgram/ml pepstatin A 10 microgram/ml leupeptin 1 microgram/ml aprotinin I have also considered adding EDTA in the breaking buffer but must remove it before an Ni-NTA binding step. If anyone has any experience in optimal inhibitor mixtures and concentrations for yeast whole cell extract (or references for the same), please let me know. Also, has anyone ever heard of the effects of various C-terminal amino acids on protease susceptibility (C-end rule?)? Thanks, Eric ________________________________________________________________________________ EARN@Portia.caltech.edu Division of Biology Abelson Lab Caltech From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!SCRI.FSU.EDU!STRELETS From: STRELETS@SCRI.FSU.EDU ("VICTOR B. STRELETS") Newsgroups: bionet.molbio.proteins Subject: Comments about secondary structure prediction server Date: 4 May 1995 14:38:56 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 92 Sender: daemon@net.bio.net Distribution: world Message-ID: <950504161716.2040bc30@scri.fsu.edu> NNTP-Posting-Host: net.bio.net Hi, Recently new server for the prediction of secondary structure was announced (bio@margit.scri.fsu.edu). In connection with lurge number of related requests, including quality of prediction, I would like to give some comment which were absent (or not detected byusers) in original announcement. 1. Method predicts secondary structure in 3-state model. States include helix, beta and coil/other ([H], [B] and [-] in reply files). Prediction of coil/other is really prediction, not just an absence of other predictions in particular parts of sequence. 2. Accuracy of the prediction was evaluated positionally, including coil/other state prediction. 3. Algorithm was learned on the content of PDB-65 (its translation to NRL3D in PIR-41). Features other than helices or beta were classified as coil/other. 4. Originally work was directed toward building (revealing) of stereochemical code in connection with formation of secondary structures, i.e. if in sequence ... in ... configuration, then secondary structure is ... in ... configuration. And such a code was revealed, of course, on some level of partial positional description. Corresponding patterns (rules) obtained in more than 95% of particular sequence patterns occuerence. Model represent secondary structure organization in general, after filtering/joining of corresponding code elements. 5. Prediction implementation - something additional to this result. It was simple idea: if it exists, why not try connect database with some prediction program and why not to expose it for public? 6. Pp.5 was probably a bad idea, because some problems immediately pop up: - mechanism of code joining, overlapping and converting code to prediction was not perfectly developed; - prediction was built as an analysis of 3 resulting weight profiles (3 states), but there are problems with weigting of different patterns in code (unexpected..) and analysis of profiles is rather simple and unsophisticated what leads to the errors in comparison even with manual profiles analysis; - thre are holes in model for some subsequences (lack of data?); - model on the server is only copy (2-4 days old) of model in work. I decided to keep server active, but people should take into account its experimental character (it was specifically mentioned in original announcement). Next week users will have in reply files FIRST three weight profiles, then automatic classification. On the profiles, sharp regions clerly indicate structures and their boundaries, even if relative ranging if profiles (due to weight problems) does not allow to draw right conclusions. Note: code revealing algorithm was organized so that any parts of code obtained with significant homology of corresponding peptides were deleted from model. So, it was orientation on the really novel proteins (or really universal code, if you like). Therefore model in application to the prediction (profiles?) could be tested even on the same learning data. Note2: mentioned by some users prevalence of helices will studied after final model building. No comment right now, except of problem with code parts weighting. Note3: any helpful comments and discussions are still welcome! Note4: if you like to use method in real work, wait one month please - some problems will be already resolved and model probably will be completed in full. And final comment (many requests, especially from disappointed people) about accuracy: test on all the bank, new proteins and specially excluded (although homology influence was blocked) from learning structures will be done after final model construction, but tests on two partial sets (excluded and set of used in buiding) shows mean positional 3-state appr. 90%. Any questions about biased sets etc. unlikely to correspond to the real situation (I checked representation in these sets), but will be resolved after final testing. And still I think that it was error to open it before finishing.. I regret, despite of some excellent replyes.. Regards, Victor B.Strelets From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!newsrelay.iastate.edu!hobbes.physics.uiowa.edu!news.uiowa.edu!news From: kpmurphy@blue.weeg.uiowa.edu (murphy kenneth p) Newsgroups: bionet.molbio.proteins Subject: Re: NO. Re: A Plea To John K! (was Re: Comment to S.Blocklehurst) Date: 4 May 1995 18:48:05 GMT Organization: University of Iowa, Iowa City, IA, USA Lines: 54 Distribution: world Message-ID: <3ob7h5$fjn@nexus.uiowa.edu> References: <950503170409.2040beb4@scri.fsu.edu> <3o9fau$9a3@news.iastate.edu> <1995May4.152111.11523@alw.nih.gov> <1995May4.171649.29042@ucl.ac.uk> NNTP-Posting-Host: red.weeg.uiowa.edu In article <1995May4.171649.29042@ucl.ac.uk>, David Jones wrote: >John Kuszewski (johnk@spasm.niddk.nih.gov) wrote: >> I will not discuss the details of the work I heard about, >> because it is not published and not my own work. > >> I will never post about unpublished work, interesting >> or not, again. > >No need to be sorry - in fact it's a shame that the nearest thing >to a half-decent scientific discussion in molbio.proteins in a year >got squelched so quickly. Of course you have a perfect right to >say as little or as much as you deem appropriate on the subject. >However, just in case there is anyone else out there who feels >like spilling the cat out of the bean bag, if George Rose has >presented work at a public meeting then it would be perfectly >reasonable for someone attending the meeting to discuss what >was presented. Let's not forget that the dissemination of ideas is >the whole point of giving a talk. Now of course if this was all >discussed privately or if the talk was say a departmental seminar - >then of course you can and must say nothing. > >Still if nothing else comes of this, it will certainly boost the >readership of George Rose's paper when it does come out! > I've been following this thread with considerable interest since I have heard Rose's talk and know John K. as well. John - I think you've handled things just fine and hope you don't take the responses personally. I don't think they were intended as flames. I agree with John that details should wait until the paper is published, which will be June or July in Proteins. I will say that the approach they're using is a build-up procedure using Monte Carlo and a simplified potential function. I was very impressed with what Rose showed and personally feel that John's estimate of ten years is too conservative. Unlike John, I'm not concerned with what the rest of us will do. To the extent that Rose and co-workers "solve" the folding problem it will tell us what the native structure will be given the sequence. It will not tell us whether that structure will be stable under specified conditions nor will it tell us how that structure is attained. These other aspects of the 'folding problem', namely stability and mechanism, leave plenty of room for continued research. Regards, Kip -- Dr. Kenneth P. Murphy e-mail: k-murphy@uiowa.edu Department of Biochemistry office: (319)335-8910 University of Iowa lab: (319)335-7936 Iowa City, IA 52242 FAX: (319)335-9570 From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!CHARLES.STANFORD.EDU!bro From: bro@CHARLES.STANFORD.EDU (David Pollock) Newsgroups: bionet.molbio.proteins Subject: RE: No! Yes! etc. Date: 4 May 1995 11:55:36 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505041854.AA12488@charles.Stanford.EDU> NNTP-Posting-Host: net.bio.net John Kuszewski (johnk@spasm.niddk.nih.gov) wrote: > I will not discuss the details of the work I heard about, > David Jones wrote: > ... it's a shame that the nearest thing >to a half-decent scientific discussion in molbio.proteins in a year >got squelched so quickly... Have to agree with Jones. I*d love to hear more of what y*all out there have heard (at public meetings) of what*s going on at the edge of the *folding problem* and what you think of it. What*s the point of the net if those of us who missed the seminar can*t hear about it? It would make a good discussion to continue, even if Kuszewski has had his say. David Pollock From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!newsserver.jvnc.net!synapse.bms.com!NewsWatcher!user From: Green_david_w.prilvms3@msmail.bms.com (David W. Green) Newsgroups: bionet.molbio.proteins Subject: Preparative CNBr cleavage Date: Thu, 04 May 1995 14:22:06 -0500 Organization: Bristol-Myers Squibb Lines: 2 Message-ID: NNTP-Posting-Host: 140.176.7.245 Is anyone familiar with preparative scale (~100 mg) methods for CNBr cleavage of proteins? From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Newsgroups: bionet.molbio.proteins,bionet.molbio.methds-reagnts Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!uchinews!ellis!scb1 From: scb1@ellis.uchicago.edu (Samuel C. Blackman) Subject: Storage of SDS/PAGE gels - a question X-Nntp-Posting-Host: midway.uchicago.edu Message-ID: Sender: news@midway.uchicago.edu (News Administrator) Reply-To: scb1@midway.uchicago.edu Organization: University of Chicago -- Academic Information Technologies Date: Thu, 4 May 1995 17:13:46 GMT Lines: 27 Xref: biosci bionet.molbio.proteins:4456 bionet.molbio.methds-reagnts:27924 Ok - I know that this is going to seem like a silly question, but please bear with me - I'm trying to settle a bet in my lab. Is it possible to store SDS/PAGE gels after running them? My advisor is curious as to whether or not you can do it without significan diffusion of the proteins. Is it possible to do so for 2-3 days? If so, what are the conditions? Finally, if you have a method for storage, is it amenable to blotting following 2-3 days of storage? This came up in our lab because we have a tech. who doesn't like to work hard and want's to run 2 gels, blot one and save the other in case her blot doesn't work. Personally, I believe that she should just do the whole thing (gel and blot) over again and not be so damn lazy. So to settle a bet amongst my fellow grad. students, I decided to ask whether or not this was possible. If someone has a suggestion or a method, I'd be more than willing to try it and post the results to the net. Please e-mail any replies, and if there is substantial interest, I'll post a summary to the group. Thanks in advance! -- Sam Blackman -- Samuel C. Blackman ! InterNet : scb1@midway.uchicago.edu MD/PhD 2/7 (Pharmacology) ! Disclaimer : Who cares what I say, I'm a student! 5513 S. Cornell Ave. #1 ! Quote : I'm not a doctor, but I play one on TV. Chicago, IL 60637-1914 ! Phone : 312/752-1082 (h) Fax: 312/996-1225 From owner-proteins@net.bio.net Wed May 03 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!daresbury!trane.uninett.no!sunic!sunic.sunet.se!seunet!news2.swip.net!doc.news.pipex.net!pipex!uknet!bcc.ac.uk!jones From: jones@bsm.bioc.ucl.ac.uk (David Jones) Subject: Re: NO. Re: A Plea To John K! (was Re: Comment to S.Blocklehurst) Message-ID: <1995May4.171649.29042@ucl.ac.uk> Date: Thu, 4 May 1995 17:16:49 GMT References: <950503170409.2040beb4@scri.fsu.edu> <1995May3.224640.20424@ucl.ac.uk> <3o9fau$9a3@news.iastate.edu> <1995May4.152111.11523@alw.nih.gov> Organization: University College London X-Newsreader: TIN [version 1.2 PL2] Lines: 34 John Kuszewski (johnk@spasm.niddk.nih.gov) wrote: > I will not discuss the details of the work I heard about, > because it is not published and not my