From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!swrinde!cam.news.pipex.net!pipex!dish.news.pipex.net!pipex!edi.news.pipex.net!pipex!sunic!sunic.sunet.se!columba.udac.uu.se!populus.slu.se!mikrob.slu.se!Jan.Svensson From: Jan.Svensson@mikrob.slu.se (Jan Svensson) Newsgroups: bionet.molbio.proteins Subject: Q Ni-NTA resin Date: Mon, 3 Jul 1995 14:37:56 GMT Organization: University of Agricultural Sciences. Dep. Microbiology Lines: 19 Message-ID: NNTP-Posting-Host: midna4.mikrob.slu.se X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Hi! I have a question about the Ni-NTA resin (qiagen/diagen). In The QIAexpressionist (booklet fronm qiagen 2 ed 1992) they state that about the Ni-NTA resin "re-use of the Ni-NTA resin..should only be performed with identical recombinant proteins..maximum of 3-5 runs per column" In the booklet there is a regenaration procedure described containing 17 steps. Isn't it possible to use the column for more than one protein as long the column is intact? Can't the regeneration be done in an easier way (less than 17 steps), just strip it with EDTA and recharge the column with NiSo4 ? Any ideas or other personal experience considering this purification system are welcome. I would prefer direct e.mail. If i get any answers i can submit em back to the group in a single posting. jan.svensson@mikrob.slu.se From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!swrinde!hookup!news.mcgill.ca!news From: Stanley Hum Newsgroups: bionet.molbio.proteins Subject: simultaneous extraction of DNA, RNA, and proteins Date: 3 Jul 1995 17:01:05 GMT Organization: McGill University Computing Centre Lines: 14 Message-ID: <3t97oh$jvp@sifon.cc.mcgill.ca> NNTP-Posting-Host: i-01.das.mcgill.ca X-Newsreader: AIR News 3.X (SPRY, Inc.) Has anyone used TRIZOL reagent (from GIBCO) to extract DNA, RNA and proteins from small tissue samples (in the order of 80 to 100 micrograms)? If not TRIZOL, is there any other method of simultaneous extractions of small amounts of tissue besides fluorometric analysis (using ethidium bromide)? I would like to use a spectrophotometer since a fluorometer is a rare item in our labs. any help would be appreciated! stan STANLEYH@BIO1.LAN.MCGILL.CA From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: wzhao80850@aol.com (WZhao80850) Newsgroups: bionet.molbio.proteins Subject: P24/CD9 western blotting antibody Date: 3 Jul 1995 16:21:52 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 6 Sender: root@newsbf02.news.aol.com Message-ID: <3t9jh0$nhs@newsbf02.news.aol.com> Reply-To: wzhao80850@aol.com (WZhao80850) NNTP-Posting-Host: newsbf02.mail.aol.com Hi there, I am looking for a western blotting antibody for P24/CD9, which is a memeber of transmembrane protein family. Any information is appreciated. Thanx in advance! Wei From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!lll-winken.llnl.gov!venus.sun.com!nntp-hub2.barrnet.net!news.Stanford.EDU!amy9.Stanford.EDU!rruss From: Ray Russ Newsgroups: bionet.virology,bionet.molbio.proteins,bionet.immunology,bionet.molbio.methds-reagnts,bionet.cellbiol Subject: gp-120 participant has questions... Date: Mon, 3 Jul 1995 13:44:55 -0700 Organization: Stanford University Lines: 29 Message-ID: References: NNTP-Posting-Host: amy9.stanford.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII In-Reply-To: Xref: biosci bionet.virology:3765 bionet.molbio.proteins:4982 bionet.immunology:4716 bionet.molbio.methds-reagnts:30398 bionet.cellbiol:2508 I was one of a group of participants in a phase II gp-120 trial which was administered by San Francisco General Hospital/UCSF several years ago. I am curious if anyone out there knows how long I will continue to test "indeterminate" on Western Blot, Elysa (sp?) and PCR. Any answers/suggestions/input would be greatly appreciated. Please keep in mind that I am for all practical pourposes a neophyte as far as immunology goes and was just a volunteer so please take it easy on the technical aspects - thanks. ______________________________________________________________________________ Ray Russ Stanford Linear Accelerator Center Operational Health Physics Department ______________________________________________________________________________ From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Newsgroups: bionet.virology,bionet.molbio.proteins,bionet.immunology,bionet.molbio.methds-reagnts,bionet.cellbiol Path: biosci!bcm!cs.utexas.edu!swrinde!emory!news-feed-1.peachnet.edu!darwin.sura.net!nih-csl!dbrml69.niaid.nih.gov!user From: Seth_Pincus@NIH.gov (Seth Pincus) Subject: Re: Epitope Mapping Kits: Chiron Mimotopes vs. Genosys Spots Kit? Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: dbrml69.niaid.nih.gov Organization: NIAID References: Date: Sun, 2 Jul 1995 15:14:07 GMT Lines: 19 Xref: biosci bionet.virology:3761 bionet.molbio.proteins:4978 bionet.immunology:4715 bionet.molbio.methds-reagnts:30384 bionet.cellbiol:2505 In article , "Ed Seijo (BIO)" wrote: > Does anyone have any experience with either of these kits or epitope > mapping in general. I would appreciate any information. > Thanks, > Ed Seijo We have published three papers describing the use of these techniques to map the human anti-HIV antibody response in fine detail. They have all been published in the Journal of Clinical Investigation 1993-4. The citations are: 91: 1987-96 93: 140-46 93: 2505-13 If these contain information germaine to your question and you want more details, then feel free to get in touch by E-mail or phone (I'm in the FASEB directory). From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!news.sprintlink.net!cam.news.pipex.net!pipex!edi.news.pipex.net!pipex!sunsite.doc.ic.ac.uk!lyra.csx.cam.ac.uk!macx26.mrc-lmb.cam.ac.uk!user From: tpg@mrc-lmb.cam.ac.uk (Tim Green) Newsgroups: bionet.molbio.proteins Subject: Re: Expression strain BL21:pLysS Date: 3 Jul 1995 11:48:01 GMT Organization: MRC Laboratory of Molecular Biology Lines: 13 Distribution: world Message-ID: References: <804549313-0-35270@blue.weeg.uiowa.edu> NNTP-Posting-Host: macx26.mrc-lmb.cam.ac.uk In article <804549313-0-35270@blue.weeg.uiowa.edu>, sfredric@blue.weeg.uiowa.edu wrote: > I would like to find out if the E.coli expression strain BL21(DE3):pLysS is > commercially available? If you have some definite info, I would appreciate > hearing from you. Thanks, Scott > rsfredri@beach.utmb.edu Novagen sell the pET system and all the associated products, including the BL21 strains. The catalogue number is 69388-1. I have no idea what the price is in America. Novagen can be contacted in the US on 800-526-7319 Tim From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!swrinde!emory!news-feed-1.peachnet.edu!darwin.sura.net!bigbird.csd.sc.edu!UNIVSCVM.CSD.SCAROLINA.EDU!N490047 From: N490047@UNIVSCVM.CSD.SCAROLINA.EDU Newsgroups: bionet.molbio.proteins Subject: Re: Osmotic shock Date: Mon, 03 Jul 95 09:27:15 EDT Organization: USC Lines: 24 Message-ID: <173D0851ES85.N490047@UNIVSCVM.CSD.SCAROLINA.EDU> References: <3sl6cr$jca@nuscc.nus.sg> NNTP-Posting-Host: univscvm.csd.sc.edu In article <3sl6cr$jca@nuscc.nus.sg> medp4003@leonis.nus.sg (Farid John Ghadessy) writes: > >Hello all, > >I've been trying to isolate a protein that I'm expressing in the >periplasm using osmotic shock (Neu and Heppel). The problem is that I >can't see any bands on an SDS gel after shocking, suggesting I'm doing >something wrong. I'm only inducing a 10ml culture to test things before I >scale up but don't seem to be able to liberate ANY proteins from the >periplasm. My questions.....are certain cell lines less "shockable" >and/or should I scale up to really see anything. > >------------------------------------------------------------------------------ Are you sure your expression system is working? I have had variable results using osmotic shock--I don't know if it is my incompetence though. A more re- producible method is to make spheroplasts with lysozyme. There is a technique for this in _CPMB_ under RNA isolation from G- bacteria. However, if your expression system is working well, you should be able to detect your protein in a crude lysis on SDS-PAGE. Let me know if you need more info. --Tom From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!swrinde!emory!news-feed-1.peachnet.edu!darwin.sura.net!bigbird.csd.sc.edu!UNIVSCVM.CSD.SCAROLINA.EDU!N490047 From: N490047@UNIVSCVM.CSD.SCAROLINA.EDU Newsgroups: bionet.molbio.proteins Subject: Re: Expression strain BL21:pLysS Date: Mon, 03 Jul 95 09:24:24 EDT Organization: USC Lines: 16 Message-ID: <173D08461S85.N490047@UNIVSCVM.CSD.SCAROLINA.EDU> References: <804549313-0-35270@blue.weeg.uiowa.edu> NNTP-Posting-Host: univscvm.csd.sc.edu In article <804549313-0-35270@blue.weeg.uiowa.edu> S. Fredricksen writes: > >I would like to find out if the E.coli expression strain BL21(DE3):pLysS is >commercially available? If you have some definite info, I would appreciate >hearing from you. Thanks, Scott You want to talk to Novagen. I believe they have an 800 number that you can get by calling 800-555-1212. If they don't have it, e-mail me and I will get it for you. --Tom >rsfredri@beach.utmb.edu > From owner-proteins@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!AARDVARK.UCS.UOKNOR.EDU!MMPAREKH From: MMPAREKH@AARDVARK.UCS.UOKNOR.EDU Newsgroups: bionet.molbio.proteins Subject: Need info on molecular approaches to study enzyme Date: 3 Jul 1995 11:09:02 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 2 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HSFNFCUNWI0008R1@aardvark.ucs.uoknor.edu> NNTP-Posting-Host: net.bio.net Hi netters, Does anyone have info on any particuar enzyme system where molecuar approahes were used to understand whether two activities are associated with one protein or two proteins? I will appreciate your help in this regard. From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!bcm!cs.utexas.edu!news.sprintlink.net!rockyd!rockyj!myersm From: myersm@rockyj.rockefeller.edu (Michael Myers) Subject: Role(s) for Acidic Domains? X-Nntp-Posting-Host: rockyj.rockefeller.edu Message-ID: Sender: notes@rockyd.rockefeller.edu (News Administrator) Organization: The Rockefeller University Date: Tue, 4 Jul 1995 23:34:16 GMT Lines: 12 What is the most recent thought on the function(s) for acidic domains in proteins? So far I have only found these in the context of transcription factors such as GCN4, Gal4, and the HMG superfamily member UBF. For the UBF proteins, these acidic domains are long runs of Asp and/or Glu -- much more dramatic than the "overall acidic" transactivation domains typified by GCN4. Has it been clearly shown that they interact with TAF proteins? I'm trying to get a foothold on a protein that shares no database homologies, but it does have a sizeable acidic domain (internal). Any comments would be appreciated. Michael Myers myersm@rockvax.rockefeller.edu From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!bcm!news.msfc.nasa.gov!sol.ctr.columbia.edu!newsxfer.itd.umich.edu!agate!library.ucla.edu!info.ucla.edu!news.bc.net!unixg.ubc.ca!hoekstra From: hoekstra@unixg.ubc.ca (Sucking Gippy) Newsgroups: bionet.molbio.proteins Subject: Recommend a lipid stain? Date: 5 Jul 1995 04:08:24 GMT Organization: University of British Columbia, Vancouver, B.C., Canada Lines: 10 Message-ID: <3td37o$4fn@nnrp.ucs.ubc.ca> NNTP-Posting-Host: interchg.ubc.ca X-Newsreader: TIN [version 1.2 PL2] Ok. So its not a protein question...but somewhat related. Has anyone ever used osmium tetroxide to stain for lipids? If so, have the results been good? And if not, can someone recommend a good stain for lipids in paraffin sections which you have had good success with? Thanks. Ken From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!agate!news.duke.edu!godot.cc.duq.edu!newsfeed.pitt.edu!gatech!howland.reston.ans.net!usenet.ins.cwru.edu!po.CWRU.Edu!rdh5 From: rdh5@po.CWRU.Edu (Robert D. Hoffman) Newsgroups: bionet.molbio.proteins Subject: Re: Recommend a lipid stain? Date: 5 Jul 1995 15:58:09 GMT Organization: Case Western Reserve University, Cleveland, OH (USA) Lines: 24 Message-ID: <3tecqh$4dm@usenet.INS.CWRU.Edu> References: <3td37o$4fn@nnrp.ucs.ubc.ca> Reply-To: rdh5@po.CWRU.Edu (Robert D. Hoffman) NNTP-Posting-Host: roo.ins.cwru.edu In a previous article, hoekstra@unixg.ubc.ca (Sucking Gippy) says: > >Ok. So its not a protein question...but somewhat related. > >Has anyone ever used osmium tetroxide to stain for lipids? If so, have >the results been good? And if not, can someone recommend a good stain for >lipids in paraffin sections which you have had good success with? > >Thanks. > >Ken > Whoa there, Ken! You are about to shoot yourself in the foot! More or less by definition, there are no lipids left in paraffin sections because they have been processed through graded alcohols and xylene before paraffin impregnation. Real cowboys do their lipid stains on air-dried frozen sections. Sudan Black and Oil Red O are personal faves. Good luck! -- Robert D. Hoffman, M.D., Ph.D.//Assistant Professor, Pathology Institute of Pathology//Case Western Reserve University 2085 Adelbert Road//Cleveland, Ohio 44106, USA rdh5@po.cwru.edu//HOFFMAN@ncifcrf.gov From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!biosci!not-for-mail From: Michael F Crowley Newsgroups: bionet.announce,bionet.cellbiol,bionet.molbio.proteins,bionet.molec-model Subject: CCL:ANNOUCE: Workshop Date: 5 Jul 1995 21:53:43 -0700 Organization: Pittsburgh Supercomputing Center, Carnegie Mellon, Pittsburgh, PA Lines: 115 Sender: biohelp@net.bio.net Approved: bionews-moderator@net.bio.net Distribution: world Message-ID: <3tfq8n$8bo@net.bio.net> References: <9507052027.AA03787@pscuxb.psc.edu> NNTP-Posting-Host: net.bio.net Xref: biosci bionet.announce:2261 bionet.cellbiol:2516 bionet.molbio.proteins:4992 bionet.molec-model:484 "METHODS OF MOLECULAR MECHANICS AND DYNAMICS OF BIOPOLYMERS" WORKSHOP Pittsburgh Supercomputing Center August 16-19, 1995 The Pittsburgh Supercomputing Center (PSC) is hosting a workshop on "Methods of Molecular Mechanics and Dynamics of Biopolymers," August 16-19, 1995. The workshop will familiarize biomedical researchers with computational methods and provide practice in applying supercomputing resources to problems of concern in molecular mechanics. Practical experience on our supercomputers will be gained in the application to: (1) the theory and practice of molecular mechanics and dynamics; (2) the development and refinement of molecular mechanics force fields; (3) the problem of conformation mapping and analysis of polypeptide structures, including the refinement of structure from measured NMR data; and (4) computation of interaction energies and free energies for protein-drug interactions and conformational thermodynamics. Workshop leaders are Dr. Charles L. Brooks III, The Scripps Research Institute and Dr. Alexander D. MacKerell Jr., University of Maryland at Baltimore. The worskhop will consist of lectures and extensive hands-on sessions. General aspects of molecular mechanics software will be discussed and a number of packages are available for use at the PSC. However, the programs CHARMM and QUANTA will be utilized most extensively in demonstrations. Hands-on sessions will be emphasized. Participants will be able to work on the examples provided or on their own experimental data. No prior supercomputing experience is necessary. This workshop is funded by a grant from the Biomedical Research Technology Program, National Center for Research Resources, National Institutes of Health. TRAVEL, MEALS AND HOTEL ACCOMMODATIONS FOR RESEARCHERS AFFILIATED WITH U.S. ACADEMIC INSTITUTIONS ARE SUPPORTED BY THIS GRANT. Enrollment is limited to 20. An application form is included. Deadline for applications is: July 10, 1995. Please direct inquires or send the following application form to blankens@psc.edu. Additional information about this workshop can be found in http://pscinfo.psc.edu/biomed/workshops95.html PITTSBURGH SUPERCOMPUTING CENTER BIOMEDICAL INITIATIVE ************************************** August 16-19, 1995 APPLICATION 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, your current 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 vita. Graduate students must have a letter of recommendation from a faculty member. Please return all application materials by July 10, 1995 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. - ------- End of Unsent Draft ------- End of Forwarded Message From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!bcm!news.msfc.nasa.gov!europa.chnt.gtegsc.com!howland.reston.ans.net!news.nic.surfnet.nl!nki.nl!nki.nl!news Newsgroups: bionet.molbio.proteins Subject: collagen Message-ID: <1995Jul4.174102.7482@nki.nl> From: coco@nki.nl (coco) Date: 4 Jul 95 17:40:58 Reply-To: Coco@nki.nl Organization: The Netherlands Cancer Institute Nntp-Posting-Host: 192.42.114.64 X-Newsreader: WinVN 0.99.5MIME-Version: 1.0Content-Type: Text/Plain; charset=ISO-8859-1Lines: 6 Lines: 6 I am looking for an antibody specific for human collagen 1 and/or collagen 3 for measuring collagen contents in supernatants of fibroblast cultures with an ELISA. Does anybody knows these kind of antibodies or such an ELISA system for collagen. From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Newsgroups: bionet.virology,bionet.molbio.proteins,bionet.immunology,bionet.molbio.methds-reagnts,bionet.cellbiol Path: biosci!ns1.faseb.org!darwin.sura.net!nih-csl!dbrml69.niaid.nih.gov!user From: Seth_Pincus@NIH.gov (Seth Pincus) Subject: Re: gp-120 participant has questions... Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: dbrml69.niaid.nih.gov Organization: NIAID References: Date: Wed, 5 Jul 1995 00:59:53 GMT Lines: 40 Xref: biosci bionet.virology:3791 bionet.molbio.proteins:4990 bionet.immunology:4736 bionet.molbio.methds-reagnts:30465 bionet.cellbiol:2514 You probably participated in one of several blinded studies. UCSF should have a study coordinator who can give you that information, plus others such as published articles resulting from the studies. But to be fair, it is highly unlikely that anyone without a detailed knowledge of your medical records can answer your questions. If your physician is not explaining these to you, get another opinion. In article , Ray Russ wrote: > I was one of a group of participants in a phase II gp-120 trial which was > administered by San Francisco General Hospital/UCSF several years ago. > > I am curious if anyone out there knows how long I will continue to test > "indeterminate" on Western Blot, Elysa (sp?) and PCR. > > Any answers/suggestions/input would be greatly appreciated. > > Please keep in mind that I am for all practical pourposes a neophyte as > far as immunology goes and was just a volunteer so please take it easy on > the technical aspects - thanks. > > > > ______________________________________________________________________________ > > Ray Russ > > Stanford Linear Accelerator Center > Operational Health Physics Department > > > > > > ______________________________________________________________________________ From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!swrinde!cs.utexas.edu!news.tamu.edu!sewild1.tamu.edu!user From: Yasha@bioch.tamu.edu (Yasha Hartberg) Newsgroups: bionet.molbio.proteins Subject: Re: Role(s) for Acidic Domains? Date: Wed, 05 Jul 1995 16:33:17 +0300 Organization: Texas A&M University Lines: 28 Message-ID: References: NNTP-Posting-Host: sewild1.tamu.edu In article , myersm@rockyj.rockefeller.edu (Michael Myers) wrote: > What is the most recent thought on the function(s) for acidic domains in > proteins? So far I have only found these in the context of transcription > factors such as GCN4, Gal4, and the HMG superfamily member UBF. For the > UBF proteins, these acidic domains are long runs of Asp and/or Glu -- > much more dramatic than the "overall acidic" transactivation domains > typified by GCN4. Has it been clearly shown that they interact with TAF > proteins? I'm trying to get a foothold on a protein that shares no > database homologies, but it does have a sizeable acidic domain > (internal). Any comments would be appreciated. I have recently been doing some literature searches into a similar question. From what I have found so far I conclude that not a lot of resaerch has been done to elucidate the roles of these or other homopolymeric repeats or amino acid rich regions in proteins. For instance, a number of developmentally regulated proteins contain homopolymeric repeats. G-box Binding Factor from Dictyostelium is nearly 50% Gln, Asn, and Ser by composition. I find these repeats remarkable and have been frustrated about the lack of information about their roles in protein structure and function. I may have some references you would find useful and would be happy to email them to you if you are interested. Yasha Hartberg Texas A&M University "The most beautiful thing in Tokyo is McDonald's." Andy Warhol From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!Germany.EU.net!EU.net!uknet!bhamcs!news.ox.ac.uk!argyll.bioch.ox.ac.uk!user From: broxholme@ (News Reader) Newsgroups: bionet.molbio.proteins Subject: Re: GST fuisons - insoluble -- HELP!!!!!! Date: 5 Jul 1995 18:10:36 GMT Organization: F1, MRC Immunochem, Biochem, Oxford Lines: 21 Message-ID: References: <3s3s1k$qe5@griffin.ccc.nottingham.ac.uk> NNTP-Posting-Host: argyll.bioch.ox.ac.uk X-Newsreader: Value-Added NewsWatcher 2.0b27+ In article , gaw1+@pitt.edu (Gary Walker) wrote: > In article <3s3s1k$qe5@griffin.ccc.nottingham.ac.uk>, Ian Goodfellow > wrote: > > > Dear All > > > > I am having quite a reall 'nightmare' trying to purify a GST fusion > > protein. > How about trying urea and/or DTT with later renaturation of the protein > for affinity chromatography. Good Luck. If you want refernces on using Sarkosyl, lowering growth temperatures and IPTG concentrations, stressing the cells osmotically during growth (the betaine-sorbitol method) or some refolding strategies, let me know. My main advice is how about changing expression system? I changed expression system (from hexahisitidine fusion to GST fusion) and my domain boundaries (putative only) and had a dramatic improvement in solubility. Chris From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Newsgroups: bionet.software,bionet.molbio.genbank,bionet.molbio.embldatabank,bionet.molbio.proteins Path: biosci!agate!howland.reston.ans.net!gatech!concert!hearst.acc.Virginia.EDU!murdoch!dayhoff.med.Virginia.EDU!wrp From: wrp@dayhoff.med.Virginia.EDU (Bill Pearson) Subject: FASTA20x1 available for DOS X-Nntp-Posting-Host: dayhoff.med.virginia.edu Message-ID: Sender: usenet@murdoch.acc.Virginia.EDU Organization: University of Virginia Date: Wed, 5 Jul 1995 13:58:33 GMT Lines: 25 Xref: biosci bionet.software:12602 bionet.molbio.genbank:2060 bionet.molbio.embldatabank:522 bionet.molbio.proteins:4987 The current version of the FASTA package has been ported to "DOS" using the Borland C++ 4.51 compiler. It is available from ftp.virginia.edu in "pub/fasta/dos". This release has not been tested extensively. Fortunately, the changes required to move the unix/mac version to DOS were modest (most changes involved changes from int to long and some work with "far" pointers). At the moment, there is no "Windows" or "Win32S" version of FASTA. If you have any problems with the program, please send me email. The FASTA package is not in the public domain. It is copyrighted by William R. Pearson and the University of Virginia. You are welcome to download the program from ftp.virginia.edu, but please do not offer it for redistribution without contacting me first. Bill Pearson -- wrp@virginia.EDU Dept. of Biochemistry #440 U. of Virginia Charlottesville, VA 22908 From owner-proteins@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!rutgers!gatech!news.sprintlink.net!howland.reston.ans.net!spool.mu.edu!bloom-beacon.mit.edu!news.bu.edu!news3.near.net!yale!news.wesleyan.edu!usenet From: Tina Guina Newsgroups: bionet.molbio.proteins Subject: Hydrophobic protein purification troubles Date: 6 Jul 1995 03:45:38 GMT Organization: Wesleyan University Lines: 73 Message-ID: <3tfm92$k5q@news.wesleyan.edu> NNTP-Posting-Host: doliver2.bio.wesleyan.edu Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="-------------------------------29401129901228" X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:bionet.molbio.proteins This is a multi-part message in MIME format. ---------------------------------29401129901228 Content-Transfer-Encoding: 7bit Content-Type: text/plain; charset=us-ascii I need help with a bit bizzare (?) problem. I am trying to overexpress and purify a small, mostly hydrophobic, most likely a membrane protein. Despite correct cloning into pET vector, I cannot see the overexpressed protein band after running the total (boiled in SDS) cell protein after SDS-PAGE. More troublesome is that the fusion of the same protein to the maltose-binding protein (MBP) is sticking to the walls of the plastic eppendorf tubes after HPLC purification. Does anyone know what I should do to avoid this, either to prevent binding to the tubes walls, or some way to solubilize it without major damage to the protein structure? Thanks a lot in advance! ---------------------------------29401129901228 Content-Transfer-Encoding: 7bit Content-Type: text/plain; charset=iso-8859-1 ---------------------------------------------------------------------------- Newsgroup: bionet.molbio.proteins * Re: ATPase activity test - Giovanni Maga (31) o Cornelius Krasel (32) * seeking an NMR job - J. Takei (23) * N-terminal blocking in mild conditions - Claudio Soares (15) * FINAL CALL FOR APPLICATIONS: IRBM practical course: frontiers of - Tim Hubbard (59) * Re: GST fuisons - insoluble -- HELP!!!!!! - Giovanni Maga (27) * Conserved Gly in Ser proteases - jlittle@biosci.arizona.edu (10) * Health and Algae - Gilly J. Strauss (8) o Neefer (17) * NAOMI Version Upgrade - Simon Brocklehurst (70) * Protein Sequence Database????? - rwhitha@eagle.cc.emory.edu (6) o Rolf Apweiler (75) * [Q] Getting structure of a particular sequence - Subhro Sen (16) * Adress of K. MATSUDA or Y.YAMAGUCHI in Japan???? - Wolfgang Sippl (6) * Rat DNA Content??? - Michele ApSimon (4) * Seeking software "O" - Peter Sibbald (8) o Gerard Kleijwegt (13) * HELP: IEF gel size - Chaan Gurer (9) o Stephen P. Driska PhD (26) * Call For Papers - R. Dana - Green Gene (7) * Cleaning DAB - Louis Hom (10) * Epitope Mapping Kits: Chiron Mimotopes vs. Genosys Spots Kit? - BIO (7) o Seth Pincus (19) + Seth Pincus (40) * Expression strain BL21:pLysS o N490047@UNIVSCVM.CSD.SCAROLINA.EDU (16) o Tim Green (13) * Need info on molecular approaches to study enzyme - MMPAREKH@AARDVARK.UCS.UOKNOR.EDU (2) * Q Ni-NTA resin - Jan Svensson (19) * Re: Role(s) for Acidic Domains? - Yasha Hartberg (28) ---------------------------------------------------------------------------- ---------------------------------29401129901228-- From owner-proteins@net.bio.net Wed Jul 05 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!news.uoregon.edu!cisco-ts5-line15.uoregon.edu!user From: tedm@darkwing.uoregon.edu (Ted M.) Newsgroups: bionet.molbio.proteins Subject: Re: Pichia recombinant proteins Date: 7 Jul 1995 04:52:11 GMT Organization: U of O Lines: 47 Message-ID: References: <3ti12p$bvi@newsbf02.news.aol.com> NNTP-Posting-Host: cisco-ts5-line15.uoregon.edu In article <3ti12p$bvi@newsbf02.news.aol.com>, bspinoza@aol.com (BSpinoza) wrote: . Consequently, we have expressed > this protein in the yeast Pichia pastoris. > 1. Are a lot of people using this system (Invitrogen)? > 2. What is the experience with improving yields? > 3. Anyone have any ideas for an interesting collaboration with this > receptor fragment? > > Kevin Harris- harrisk@uthscsa.edu Kevin, The Pichia system was developed by Philips Petroleum, I believe, as a single cell food system. They developed these incredibly high yield fermentations and then it kind of dead ended. Then recombinant proteins appeared and they knew enough Pichia genetics to create an expression system. Originally they offered the expression system as a package deal for $10,000 (or was it $30,000?) where they would sell you the strain, the vectors and then help you troubleshoot expression. There were few takers. While the cell density was very high, similar S. Cerv fermentations are possible with much better systems. There was no guarantee it would work for your system, either. I think the few takers were companies with big wallets and small brains, molecular biologically speaking. Later they gave a license to Invitrogen and the system is reborn. The main advantage of the system has always been the possibility of very high density growth and decent secreted protein. This advantage is best realized with fermentation. Smaller scale cultures are possibly no better than S. cerv. systems, IMHO. Keep in mind that similar sized glycosylation does not mean native glycosylation; my experience is that these homologs tend to be cleared much faster from the blood, albeit not as fast as ³naked² bacterial proteins, or ³aberrantly glycosylated² insect proteins. The typical way to improve Pichia yields is to produce huge cell masses and then induce via the methanol promoter in a two stage fermentation. There should be some ref. (included?) from th earlier Philips Pet. work. Often cystein bridges are the undoing of these fungal systems, renaturation can help, possibly with disulfide isomerase. Good luck! Ted Michelini Institute of Molecular Biology University of Oregon Tedm@darkwing.uoregon.edu From owner-proteins@net.bio.net Wed Jul 05 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: bspinoza@aol.com (BSpinoza) Newsgroups: bionet.molbio.proteins Subject: Pichia recombinant proteins Date: 6 Jul 1995 21:02:17 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 16 Sender: root@newsbf02.news.aol.com Message-ID: <3ti12p$bvi@newsbf02.news.aol.com> Reply-To: bspinoza@aol.com (BSpinoza) NNTP-Posting-Host: newsbf02.mail.aol.com I have been studying the human erythropoietin receptor extracellular domain as a bacterially expressed GST fusion protein. I became unsatisfied with the activity of this recombinant protein- only 1% of the soluble protein bound ligand with the expected high affinity, even after tedious affinity purification. Consequently, we have expressed this protein in the yeast Pichia pastoris. It seems to have a much higher specific activity than the bacterially expressed protein (still working on how much...) and is appropriately glycosylated (by molecular weight). Unfortunately the yields are, to say the least, disappointing. I have three questions: 1. Are a lot of people using this system (Invitrogen)? 2. What is the experience with improving yields? 3. Anyone have any ideas for an interesting collaboration with this receptor fragment? Kevin Harris- harrisk@uthscsa.edu From owner-proteins@net.bio.net Wed Jul 05 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!ix.netcom.com!netnews From: pmcatee@ix.netcom.com (Patrick McAtee ) Newsgroups: bionet.molbio.proteins Subject: COOH sequencing, ATPase, IEF.... Date: 6 Jul 1995 22:37:12 GMT Organization: Netcom Lines: 12 Distribution: world Message-ID: <3thoio$elj@ixnews5.ix.netcom.com> NNTP-Posting-Host: ix-pa3-24.ix.netcom.com I'm reading the ATPase posts, could somebody provide me with a reference for ATPase assay using PEI/TLC ? Does anybody know of a rapid COOH terminal (CyP) sequencing facility that can get me good results, quick ? IEF/Western blot-my westerns are coming out fuzzy ? C. Patrick McAtee, Ph.D GeneLabs Technologies FAX (415) 368-0709 From owner-proteins@net.bio.net Wed Jul 05 23:00:00 1995 Path: biosci!NMSU.EDU!hroychow From: hroychow@NMSU.EDU (Hiranya Roychowdhury) Newsgroups: bionet.molbio.proteins Subject: Re: Hydrophobic protein purification troubles Date: 6 Jul 1995 14:03:47 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 43 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <3tfm92$k5q@news.wesleyan.edu> NNTP-Posting-Host: net.bio.net On 6 Jul 1995, Tina Guina wrote: > I need help with a bit bizzare (?) problem. > I am trying to overexpress and purify > a small, mostly hydrophobic, most likely > a membrane protein. > Despite correct cloning into pET vector, > I cannot see the overexpressed protein band > after running the total (boiled in SDS) cell If it is hydrophobic and membrane associated, may be you are losing it with the bacterial pellet. Another possibility is aberrant mobility due to its hydrophobic nature: hydrophobic proteins will tend to bind SDS at a higher ratio compared to hydrophilic ones, thereby migrating slightly faster on a SDS-PAGE. Thus, you may be expecting a 40kDa band while the polypetide is at 35kDa (just an example) > protein after SDS-PAGE. > More troublesome is that the fusion of the > same protein to the maltose-binding protein > (MBP) is sticking to the walls of the plastic > eppendorf tubes after HPLC purification. More pure a hydrophobic protein the more will be its propencity to bind to plastic (hydrophobic) surfaces. Try siliconizing. Does anyone know what I should do to avoid this, > either to prevent binding to the tubes walls, or > some way to solubilize it without major damage to > the protein structure? > > Thanks a lot in advance! > > > >>>>>>>>>>>>>>>>>>>>>>>>>>> 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 Wed Jul 05 23:00:00 1995 Path: biosci!galaxy.ucr.edu!ratatosk.yggdrasil.com!nntp-sc.barrnet.net!nntp-hub2.barrnet.net!news3.near.net!yale!news.wesleyan.edu!usenet From: Tina Guina Newsgroups: bionet.molbio.proteins Subject: Hydrophobic protein purification Date: 6 Jul 1995 16:55:50 GMT Organization: Wesleyan University Lines: 12 Message-ID: <3th4im$41j@news.wesleyan.edu> NNTP-Posting-Host: doliver2.bio.wesleyan.edu 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:bionet.molbio.proteins I have problems purifying the fusion of a small, hydrophobic, most likely hydrophobic protein to maltose-binding protein (MBP). Fusion protein sticks to the walls of the plastic (eppendorf) tubes. Does anybody know how to avoid this problem and how to remove the protein from the tube walls without destruction of the protein structure? Thanks a lot! From owner-proteins@net.bio.net Wed Jul 05 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!agate!news.mindlink.net!vanbc.wimsey.com!pm056.bby.wis.net!gordonr From: gordonr@wimsey.com (Gordon Robertson) Newsgroups: bionet.molbio.proteins Subject: Removing lipids from enzymes in aqueous media Date: Thu, 6 Jul 1995 20:46:38 Organization: PAPRICAN Lines: 17 Message-ID: NNTP-Posting-Host: pm056.bby.wis.net Keywords: lipids, lipase, enzyme, purification X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Can anyone help with a colleague's problem? He is trying to purify an extracellular lipase produced by a fungus in aqueous media containing high concentrations of peptone and olive oil. It appears that part of the lipase is bound to the oil phase. When he tries to precipitate the enzyme with ammonium sulphate, at concentrations of salt below 70% he recovers < 40% of the activity. However, at high concentration, 80-85% salt, he gets no better yield, as some of the protein stays in the upper phase with the oil, and does not enter the pellet. He has tried removing the oil with immiscible solvents, but this is to harsh, and lowers the enzyme activity. Can anyone suggest an approach to this problem? Thank you in advance for your time in this. From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!swrinde!hookup!news.mcgill.ca!news From: Elida@parasit.lan.mcgill.ca Newsgroups: bionet.molbio.proteins Subject: Triton-X 100 effect on protein concentration determination Date: 6 Jul 1995 20:08:26 GMT Organization: McGill University Computing Centre Lines: 5 Message-ID: <3thfrq$6t3@sifon.cc.mcgill.ca> NNTP-Posting-Host: swift.parasitology.mcgill.ca X-Newsreader: AIR News 3.X (SPRY, Inc.) Effect of Triton X-100 on protein concentration determination. Has anyone in this net ever evaluated the effect of Triton-X 100 present in protein sample when measuring protein concentration by the BioRad assay? If you did, please tell me what happened. Elida@parasit.lan.mcgill.ca From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!ns1.faseb.org!darwin.sura.net!fconvx.ncifcrf.gov!mack From: mack@ncifcrf.gov (Joe Mack) Subject: Re: Need Help in Purification Message-ID: Organization: Frederick Cancer Research and Development Center References: <3tjudv$qp8@newsbf02.news.aol.com> Date: Fri, 7 Jul 1995 19:15:52 GMT Lines: 28 In article <3tjudv$qp8@newsbf02.news.aol.com> jt31@aol.com (JT31) writes: >I am currently in the process of purifying an NADP Dehydrogenase from E. >coli. My first procedure is Ammonium Sulfate fractionation....My protein >precipitates at approximately 30% salt conc., however the activity is much >lower (about a 35% loss). This is normal in my experience. Even after desalting through a Sephadex G-10 >column there is no increase in activity. Normally after that I put the >semi-crude extract through a Ion exchange process using a DEAE Celluolose >resin. Again the procedure works out find, the enzyme is succesfully >eluted (Using 300mM Tris HCl pH 7.2), However More activity is lost. >Finally After running the extract through a Cibacron Blue resin, and >eluting with approx 1.2M NaCl, the porotein is successfully retrieved but >again more activity is lost! All procedures are performed in a cold room >as well. Does anyone have any hints, tips, or tricks for keeping this >enzyme active??? I would greatly appreciate the Info. > >Dominic Ricciardi Wouldn't worry about it - recovering 10% of activity after 3 steps happens a lot. As you get more experience with the protein, you might get better yields, but for now as long as the enzyme is being purified, then there may not be much you can do. Joe Mack mack@ncifcrf.gov From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!Norway.EU.net!EU.net!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: jt31@aol.com (JT31) Newsgroups: bionet.molbio.proteins Subject: Looking for an NADP Dehydrogenase Date: 7 Jul 1995 14:17:33 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 7 Sender: root@newsbf02.news.aol.com Message-ID: <3tjtnt$qkh@newsbf02.news.aol.com> Reply-To: jt31@aol.com (JT31) NNTP-Posting-Host: newsbf02.mail.aol.com I would appreciate it if anyone has come across an NADP Dehydrogenase that is fairly alkali tolernant. I am in the process of Isolating a protein with these characteristics in E. coli. Would appreciate any info on an enzyme with similar characteristics found in E. coli or in any other organism. Dominic Ricciardi From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!news.uoregon.edu!kaiwan.kaiwan.com!NewsWatcher!user From: QIAGEN@kaiwan.com (QIAGEN) Newsgroups: bionet.molbio.proteins Subject: Re: Q Ni-NTA resin Followup-To: bionet.molbio.proteins Date: 7 Jul 1995 16:22:45 GMT Organization: Kaiwan Lines: 45 Distribution: world Message-ID: NNTP-Posting-Host: kaiwan009.kaiwan.com In Message-ID: Jan.Svensson@mikrob.slu.se (Jan Svensson) wrote: >I have a question about the Ni-NTA resin (qiagen/diagen). >In The QIAexpressionist (booklet fronm qiagen 2 ed 1992) they state that about >the Ni-NTA resin "re-use of the Ni-NTA resin..should only be performed with >identical recombinant proteins..maximum of 3-5 runs per column" >In the booklet there is a regenaration procedure described containing 17 steps. >Isn't it possible to use the column for more than one protein as long the >column is intact? >Can't the regeneration be done in an easier way (less than 17 steps), just >strip it with EDTA and recharge the column with NiSo4 ? Hi Jan, Ni-NTA resin can probably be reused after stripping the column with EDTA and recharging it with Ni ions. However, this will give you no guarantee that you have removed all the residual proteins and you risk contamination of your next prep. Also, the resin properties will progressively deteriorate as the level of residual contamination increases. The long washing procedure we describe is tedious, we know, but it is the only way to be absolutely sure you have removed all contaminants (both protein and non-protein) that may be interacting with the resin. Most users find that it is safer, more convenient, and more cost-effective (unless you are using extremely high resin volumes) to use fresh Ni-NTA resin where possible. Also, you should note that Ni-NTA resin will bind 5Š10 mg of protein per ml of resin, and that the purification procedure is more efficient if you use less resin than you actually need, and overload it somewhat. The QIAGEN Technical Service Department has extensive references for this system, and can help you optimize your purification procedure by using less Ni-NTA resin. QIAGEN Technical Service Department 800-426-8157 -- QIAGEN 800-362-7737 Temporary Email Address qiagen@kaiwan.com From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!europa.chnt.gtegsc.com!newsxfer.itd.umich.edu!jobone!lynx.unm.edu!bubba.NMSU.Edu!dkim From: dkim@nmsu.edu (Daniel Kim) Newsgroups: bionet.molbio.proteins Subject: Re: Triton-X 100 effect on protein concentration determination Date: 7 Jul 1995 17:56:41 GMT Organization: New Mexico State University, Las Cruces, NM Lines: 31 Sender: dkim@nmsu.edu Distribution: world Message-ID: <3tjsgp$818@bubba.NMSU.Edu> References: <9507071348.AA04629@bnlux1.bnl.gov.bnl.gov> NNTP-Posting-Host: verdi.nmsu.edu I have been reading this thread, and thought this might be relevant: I am using a protein determination kit called dotMETRIC. It is a spot-blot kind of assay, in which the diameter of the spot on a gel is proportional to the total amount of protein. Thus, a single spot of 1 microliter of unknown protein solution can be used to get a reasonably accurate measure of total protein. The manufacturers claim that the assay is unaffected by detergents, and does not require a protein standard curve. It gives the same result regardless of the individual protein characteristics. I am new at this protein game (Proteins have always been the stuff I throw away in the phenol phase), so I thought the kit would be worth a try. It does seem to be very easy and consistent. I have made no formal comparisons with other assay methods, however. Has anyone else tried this kit and made a comparison? If interested, here is the address, etc for the manufacturer: Geno Technology, Inc. 3803 Washington Blvd. St. Louis, MO 63108 USA (314) 534-0075 FAX (314) 534-1883 Daniel Kim (no affiliation with the company.) From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!news.moneng.mei.com!news.ecn.bgu.edu!psuvax1!news.eecs.nwu.edu!newsfeed.acns.nwu.edu!uicvm.uic.edu!u12201 Organization: University of Illinois at Chicago, ADN Computer Center Date: Fri, 7 Jul 1995 13:14:46 CDT From: Message-ID: <95188.131447U12201@uicvm.uic.edu> Newsgroups: bionet.molbio.proteins Subject: Re: Triton-X 100 effect on protein concentration determination References: <3thfrq$6t3@sifon.cc.mcgill.ca> Lines: 8 The Pierce BCA assay works like a charm even in the presence of very high conc. of TX100. Any Coomassie type assay (Bradford) will give high backgrounds values with TX100. No way around it!! I personally prefer BCA for its versatility and flexibility of the protocol (you can "tune" the sensitivity by varying the time and the temp.) KeldS@uic.edu From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!news.uoregon.edu!europa.chnt.gtegsc.com!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: jt31@aol.com (JT31) Newsgroups: bionet.molbio.proteins Subject: Need Help in Purification Date: 7 Jul 1995 14:29:19 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 15 Sender: root@newsbf02.news.aol.com Message-ID: <3tjudv$qp8@newsbf02.news.aol.com> Reply-To: jt31@aol.com (JT31) NNTP-Posting-Host: newsbf02.mail.aol.com I am currently in the process of purifying an NADP Dehydrogenase from E. coli. My first procedure is Ammonium Sulfate fractionation....My protein precipitates at approximately 30% salt conc., however the activity is much lower (about a 35% loss). Even after desalting through a Sephadex G-10 column there is no increase in activity. Normally after that I put the semi-crude extract through a Ion exchange process using a DEAE Celluolose resin. Again the procedure works out find, the enzyme is succesfully eluted (Using 300mM Tris HCl pH 7.2), However More activity is lost. Finally After running the extract through a Cibacron Blue resin, and eluting with approx 1.2M NaCl, the porotein is successfully retrieved but again more activity is lost! All procedures are performed in a cold room as well. Does anyone have any hints, tips, or tricks for keeping this enzyme active??? I would greatly appreciate the Info. Dominic Ricciardi From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!agate!howland.reston.ans.net!news.sprintlink.net!dish.news.pipex.net!pipex!oleane!jussieu.fr!centre.univ-orleans.fr!univ-lyon1.fr!news-rocq.inria.fr!irisa.fr!news.univ-rennes1.fr!news.univ-brest.fr!molene!news From: cladrat@ifremer.fr (Christine Ladrat, Ifremer Brest PDG-DRO-EP) Subject: Need info on E. coli lacking ADH activity Message-ID: <1995Jul7.121231.9360@molene.ifremer.fr> Sender: news@molene.ifremer.fr Organization: ifremer X-Newsreader: WinVN 0.92.5 Date: Fri, 7 Jul 95 12:12:31 GMT Lines: 4 I am trying to clone a gene encoding a thermostable alcohol dehydrogenase. In order to screen by expressed activity, I would like to find a host strain lacking ADH. Do you something about that ? Thanks for reply. C. Ladrat From owner-proteins@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!BNLUX1.BNL.GOV!berges From: berges@BNLUX1.BNL.GOV (John A. Berges) Newsgroups: bionet.molbio.proteins Subject: Re: Triton-X 100 effect on protein concentration determination Date: 7 Jul 1995 06:49:55 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 35 Sender: daemon@net.bio.net Distribution: world Message-ID: <9507071348.AA04629@bnlux1.bnl.gov.bnl.gov> NNTP-Posting-Host: net.bio.net >>>Effect of Triton X-100 on protein concentration determination. >>>Has anyone in this net ever evaluated the effect of Triton-X 100 present in >>protein sample when measuring protein >>>concentration by the BioRad assay? If you did, please tell me what happened. >>>Elida@parasit.lan.mcgill.ca >>> The effect of detergents on the Bradford (i.e. Biorad) protein assay is well documented...it really messes things up. The absorbance of all your samples will go sky-high. Biorad says up to about 0.1% Triton can be compensated for by including the detergent in standards, but this leads to samples that change very quickly in absorbance and precipitate within an hour or so. Also, there are marked non-linearities in standard curves with Triton. Biorad also has a detergent compatible assay, based on the Lowry procedure. Bicinchoninic acid-based assays also work well with detergents; see kits by Sigma, Pierce and others. These assays won't work in the presence of reducing agents (e.g. DTT or mercaptoethanol). Recently, Geno Technology has come out with a "dotMetric" assay that makes some big promises in advertising (although the accompanying literature qualifies many of these). It is supposed to overcome just about any commoninterference. I haven't tried it yet...anyone else out ther have any experience with it? Cheers. ________________________________________________________________ _/_/_/ _/ _/ _/ Dr. John A. Berges _/ _/ _/ _/ _/ Oceanic and Atmospheric _/ _/ _/_/ _/ _/ Sciences Divison _/_/_/ _/ _/ _/ _/ Brookhaven National Lab _/ _/ _/ _/ _/ _/ Upton, NY 11973 _/ _/ _/ _/_/ _/ Phone: 516-282-4077 _/_/_/_/ _/ _/ _/_/_/_/ Email: berges@BNLUX1.BNL.GOV ________________________________________________________________ From owner-proteins@net.bio.net Fri Jul 07 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!nntp.gmd.de!news.rwth-aachen.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!news.belwue.de!News.Uni-Marburg.DE!news.th-darmstadt.de!fauern!lrz-muenchen.de!ipp-garching.mpg.de!genmac8.biochem.mpg.de!user From: spang@vms.biochem.mpg.de (Anne Spang) Newsgroups: bionet.molbio.proteins Subject: Re: Triton-X 100 effect on protein concentration determination Date: 8 Jul 1995 09:40:59 GMT Organization: Rechenzentrum der Max-Planck-Gesellschaft in Garching Lines: 8 Message-ID: References: <3thfrq$6t3@sifon.cc.mcgill.ca> NNTP-Posting-Host: genmac8.biochem.mpg.de Hi! Triton X-100 messes up your Bradford assay! In the presence of detergent in our sample, we usually precipitate the sample with TCA in a dotblot apparatus on nitrocellulose, then stain with amido black. After elution teh samples can be read in an ELISA reader or a normal photometer. Hope this helps Anne From owner-proteins@net.bio.net Fri Jul 07 23:00:00 1995 Path: biosci!agate!newsxfer.itd.umich.edu!europa.chnt.gtegsc.com!news.sprintlink.net!dish.news.pipex.net!pipex!uknet!daresbury!s-crim1!mbkdj From: mbkdj@s-crim1.dl.ac.uk (K.D. James) Newsgroups: bionet.molbio.proteins Subject: Re: Need Help in Purification Date: 8 Jul 1995 18:26:53 GMT Organization: SERC Daresbury Lab, Warrington, U.K. Lines: 45 Distribution: bionet Message-ID: <3tmild$q87@mserv1.dl.ac.uk> References: <3tjudv$qp8@newsbf02.news.aol.com> NNTP-Posting-Host: s-crim1.dl.ac.uk X-Newsreader: TIN [version 1.2 PL2] JT31 (jt31@aol.com) wrote: : I am currently in the process of purifying an NADP Dehydrogenase from E. : coli. My first procedure is Ammonium Sulfate fractionation....My protein : precipitates at approximately 30% salt conc., however the activity is much : lower (about a 35% loss). Even after desalting through a Sephadex G-10 : column there is no increase in activity. Normally after that I put the : semi-crude extract through a Ion exchange process using a DEAE Celluolose : resin. Again the procedure works out find, the enzyme is succesfully : eluted (Using 300mM Tris HCl pH 7.2), However More activity is lost. : Finally After running the extract through a Cibacron Blue resin, and : eluting with approx 1.2M NaCl, the porotein is successfully retrieved but : again more activity is lost! All procedures are performed in a cold room : as well. Does anyone have any hints, tips, or tricks for keeping this : enzyme active??? I would greatly appreciate the Info. : Dominic Ricciardi I have been purifying NAD dehyrogenases from Gram negatives using dye-ligand columns (inc Cibacron blue) and DEAE cellulose. The following questions spring to mind: Is your loss of activity due to poor stability or poor recovery from the columns? My enzymes can suffer from the former, so adding 2mM DTT or 20% glycerol (depends on the enzyme) can help retain activity. As far as I'm aware, Cibacron Blue is good for both NAD and NADP linked enzymes, but have you tried Procion Red? I'm sure this has a high affinity for NADP linked types. Also, if you have to raise the NaCl concentration to 1.2M to recover your enzyme there is the possibility of using a lower concentration to elute unwanted protein while leaving yours bound, followed by a more biospecific elution with NAD/P. Of course, cofactor elutions may well be something you've already tried. (You can get Procion Red HE-3B as Matrex Gel Red A from Amicon.) Currently I use a Cibacron Blue column as a first step, passing cell extracts straight on to it (okay if your matrix can be regenerated harshly- I use a 6% cross-linked agarose bead type which can be cleaned with 8M urea). This followed by a DEAE Sephacel step, eluting with an NaCl gradient. I'm also always interested in protein purification tips... Keith James (No connection with Amicon etc.) From owner-proteins@net.bio.net Sat Jul 08 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!gatech!howland.reston.ans.net!Germany.EU.net!zib-berlin.de!informatik.tu-muenchen.de!lrz-muenchen.de!news From: Wolfgang Schechinger Newsgroups: bionet.molbio.proteins Subject: Re: Hydrophobic protein purification troubles Date: 9 Jul 1995 21:08:41 GMT Organization: Leibniz-Rechenzentrum, Muenchen (Germany) Lines: 14 Distribution: world Message-ID: <3tpggp$rr0@sparcserver.lrz-muenchen.de> References: <3tfm92$k5q@news.wesleyan.edu> NNTP-Posting-Host: u7k0201.ppp.lrz-muenchen.de Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: hroychow@NMSU.EDU >> More troublesome is that the fusion of the >> same protein to the maltose-binding protein >> (MBP) is sticking to the walls of the plastic >> eppendorf tubes after HPLC purification. > >More pure a hydrophobic protein the more will be its propencity to bind >to plastic (hydrophobic) surfaces. Try siliconizing. > Doesn't siliconizing meking the surface even more hydrophobic? I would try a glass tube instead. Wolfgang From owner-proteins@net.bio.net Sat Jul 08 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!news.sprintlink.net!gatech!bloom-beacon.mit.edu!spool.mu.edu!torn!ccshst05.cs.uoguelph.ca!ccshst01.cs.uoguelph.ca!chouse From: chouse@uoguelph.ca (Chris J House) Newsgroups: bionet.molbio.proteins Subject: Lactobacil Date: 8 Jul 1995 21:15:47 GMT Organization: University of Guelph Lines: 6 Message-ID: <3tmsi3$per@ccshst05.cs.uoguelph.ca> NNTP-Posting-Host: ccshst01.cs.uoguelph.ca X-Newsreader: TIN [version 1.2 PL2] Does anyone have current info pertaining to the protein channels and their efficiancy in L.brevis? From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!EU.net!Austria.EU.net!newsfeed.ACO.net!news.univie.ac.at!helix.mdy.univie.ac.at!hs From: hs@helix.mdy.univie.ac.at (Hellfried Schreiber) Newsgroups: bionet.molbio.proteins Subject: INT.CONF.on MOLEC.STRUCT.BIOL. - Vienna 95 Date: 10 Jul 1995 12:02:52 GMT Organization: Inst. for Theoretical Chemistry / Univ. of Vienna Lines: 1018 Distribution: world Message-ID: <3tr4tc$4g7e@ftp.univie.ac.at> NNTP-Posting-Host: helix.mdy.univie.ac.at There is a opportunity to sumbit late-braking abstracts for presentation at the >> INTERNATIONAL CONFERENCE ON MOLECULAR STRUCTURAL BIOLOGY << Vienna (Austria), September 17-20, 1995 ================================================================================ ================= the DEADLINE is the 7th of AUGUST, 1995.====================== ================================================================================ For details see below: -------------------------------------------------------------------------------- >> INTERNATIONAL CONFERENCE ON MOLECULAR STRUCTURAL BIOLOGY << Organized by the Austrian Chemical Society Vienna (Austria), September 17-20, 1995 TOPICS: 1) The Impact of Molecular Biology on Structural Biology 2) Biomolecular Structure Determination a) X-Ray Diffraction b) NMR Spectroscopy 3) Dynamics and Function of Biomolecules 4) Computational Methods 5) Protein Engineering and Design VIENNA: The organisers would like to invite you to a new conference in the rapidely expanding field of molecular structural biology. The conference will take place in Vienna, home of waltz, the capital of a fomrer great empire, one of the three centres of the United Nations and a leading host city in the world for important meetings and conferences. Scientific topics to be presented at this conference will highlight the state-of-the-art in biomolecular structure determination in its context of molecular biology, medicine and pharmacology. Scientists from both basic and industrial research are highly welcome to present their results to an interested and enthusiastic audience. In addition to the conference itself, you could take time to explore the wonderful city of Vienna, perhaps wandering through the elegant parks in their autumnal colours before soaking up the rich atmosphere of the renowned Viennese cafes. A reception with the mayor in the historic Rathaus at the end of the first day of presentations will provide a taste of the rich Viennese culture waiting to be discovered. ORGANISING COMMITTEE: A.Kungl, P.Andrews, B.Habermann, H.Schreiber, A.Schrenk SCIENTIFIC COMMITTEE: Chairman: P.Schuster M.Breitenbach, C.Kratky, P.Andrews, H.Schreiber A.Kungl IN COOPERATION WITH Austrian Academy of Sciences Federal Ministry of Economy Federal Ministry of Science and Research SPONSORED BY Wirtschaftskammer Oesterreich SCIENTIFIC PROGRAMME: The scientific programme is designed to cover a broad range of disciplines in molecular structural biology. Topics will be presented in the form of plenary lectures, posters, discussions and exhibitions. Sunday, (17.9) ----------------------------------------------------------------------------- Registration Welcome Cocktail Honory Lecture by H.Neurath Monday, (18.9) ----------------------------------------------------------------------------- 1. The impact of Molecular Biology on Structural Biology C.Cantor (USA) "Application of Streptavidin and Genetically Engineered Variants" T.Blundell (GB) "Protein Super Families: Genome Analyses and Drug Discovery" 2. Biomolecular Structure Determination a) X-Ray Crystallography R.Huber (D) "Proteolytic Enzymes and Their Natural Inhibitors - New Functions and New Structures and No End in Sight" M.Walkinshaw (CH) "Structures and Biological Implications of Series of Cyclophilin-Cyclosporin Crystal Complexes" I.Schlichting (D) "Ligand Binding in Heme Proteins Studied by Kinetic X-Ray Crystallography" Tuesday (19.9.) ----------------------------------------------------------------------------- b) NMR Spectroscopy O.Jardetzky (USA) "Flexibility and Function in a DNA Binding Protein - the Trp-Repressor of E.Coli" P.Roesch (D) "The Structure of Iron-Sulfur Proteins in Solution" C.Dobson (GB) "The Structural Basis of Protein Folding" 3. Dynamics and Function of Biomolecules J.Lakowicz (USA) "Fluorescence Spectroscopic Studies of the Structure and Dynamics of Biomolecules" W.Kuehlbrandt (D) "Structure Determination of Membrane Proteins by High-Resolution Electron Microscopy" H.Frauenfelder (USA) "Dynamics and Function of Biomolecules" Wednesday (20.9.) ----------------------------------------------------------------------------- 4. Computational Methods M.Karplus (USA) "Simulations of Protein Folding From the Native to the Denatured State and Back Again" J.Skolnick (USA) "A Hierarchial Approach to the Prediction of Protein Structures" H.-J.Boehm (D) "New Computational Tools for Structure Based Drug Design" 5. Protein Engineering and Design A.Fersht (GB) "Pathways of Protein Folding" L.Presta (USA) "Protein Engineering of Immunoglobulins and Immunoglobulin-Like Domains" M.Mutter (CH) "De Novo Protein Design" CALL FOR POSTERS ----------------- All those intending to participate in the conference are invited to submit posters presenting original work. Abstracts prepared according to the instructions given below should arrive NOT LATER THAN AUGUST 7th, 1995. The authors will be informed about the provisional acceptance in July, 1995. The presentation of the posters will be finally approved and thus the contribution included in the Book of Abstracts when at least one of the registers before August 1st, 1995. Contributors of outstanding posters will be chosen by the scientific committee to give a 20 minute lecture. Abstracts: Camera-ready single-page abstacts of posters should start with the title of the contribution followed by the author(s) name(s), affiliation(s) and address(es). Abstracts must be typed within an area 16cm wide and 24cm long, centred on a sheet of (about) A4 size (210mm x 297mm). The chosen topic must be assigned by stating its title (see list of topics) in the top right margin. The abstracts should be sent unfolded to the conference secreteriat. Proceedings: Invited lectures presented at the conference will be published in book form. All authors will receive in due course instructions for preparing the manuscript (for type-setting, not camera-ready). The deadline for submitting manuscripts is June 30, 1995. Posters: The size of the posters board is 0.9m (width) x 1.5m (height). All posters will be numbered according to their order in the book abstracts. Thumbtacks, tape and markers will be available in the poster area. Head of Scientific Committee: P.Schuster GENERAL INFORMATION: -------------------- EXHIBITON An exhibition of instruments, accessories, software, literature and other items is planned. Companies interested in displaying their products are kindly requested to contact the conference secretariat. INDUSTRIAL SATELLITE MEETING A half day industrial symposium is planned. Those companies who are interested in presenting up-to-date results obtained on their products in the form of 20 minute lectures are kindly requested to contact the conference secretariat. SOCIAL EVENTS The conference will be opened by the welcome cocktail and buffet in the rooms of the Austrian Bundeswirtschaftskammer on Sunday evening, September 17. On Monday, September 18, the mayor of Vienna will invite the conference participants to the historical Viennese Rathaus. A piano concert will take place on Tuesday, September 19, in the hall of the Austrian Gewerbeverein. A farewell evening at a typical Austrian wine tavern is planned for the evening of Wednesday, September 20. Activities for accompanying persons will include cultural events and visits to places of general interest. REGISTRATION To enjoy the early rates, please complete the enclosed Registration Form and send it to the conference secreteriat before August 1, 1995. Registration Fees: before August 1 after August 1 Regular Participant 4000 ATS 4500 ATS GOeCH Member 3500 ATS 4000 ATS Student 2000 ATS 2300 ATS Accompanying Persons 500 ATS 700 ATS The registration desk will be open from 2 p.m. on Sunday, September 17. The registration fee includes the Book of Abstracts, the welcome cocktail and buffet, the invitation to the Viennese Rathaus, a ticket for the piano concert, coffee breaks and tram tickets. ! Meals and accomodation are not included in the registration fee! Tickets for lunch will be available for 190ATS/meal/person (see Registration Form). For accomodation please fill in and return the enclosed flyer of the travel agency MONDIAL CONGRESS to the address given. Remittance of Fees: Remittance must be made in Austrian Schillings (ATS) payalbe to the Gesellschaft Oesterreichischer Chemiker, Abeitsgruppe Biophysikalische Chemie, account number 0043-19265/o4 at Bank Creditanstalt, banking code 11000. Payment is also accepted by sending (Euro-)Cheque to the conference secreteriat or by filling in the credit card form (Visa, Euro/Mastercard, Dinersclub and American Express are accepted). All charges due to bank transfer have to be paid by the sender. The sender's name and address have to be marked clearly on every remittance. Canellation: Applications may be cancelled up to August 15, in which case 85% refund of fees already paid will be made. It will not be possible to offer any refunds if cancellations are received after that date. Key dates: --------- June 30 deadline for submitting poster abstracts and manuscripts August 1 deadline for early registration fee August 15 deadline for cancellation Location: --------- Wirtschaftskammer Oesterreich (Julius Raab Saal) Wiedner Hauptstrasse 63 A-1040 Vienna Conference Secretariat: ---------------------- Gesellschaft Oesterreichischer Chemiker AG Biophysikalische Chemie Dr. Andreas Kungl Nibelungengasse 11 A-1010 Wien, Austria Tel.: (43) 1 587249 FAX.: (43) 1 587966 e-mail: msb95@helix.mdy.univie.ac.at _______________________________________________________________________________ REGISTRATION FORM: Surname........................First Name .....................Title........... Institution/Company............................................................ Departement.................................................................... Street....................................Town................................. Country...................................Post Code............................ Tel.Nr....................................Fax Nr............................... e-mail......................................................................... I am a(n) ...Abstract Submitter ...Invited Speaker ...Regular Participant ...GOECH-Member ...Student Title of Poster/Lecture........................................................ ............................................................................... ...I will require ...Mo ...Tue ...Wed lunch tickets (190 ATS/meal/person) (Payment should accompany registration) Method of Payment: ...Bank Transer to Bank Creditanstalt account number 0043-19265/04, bank code 11000 ...(Euro-)Cheque payalbe to Gesellschaft Oesterreichischer Chemiker, 'International Conference on Molecular Structural Biology' ...Credit Card ...Visa ...Euro/Master ...American Express ...Diners Club Card Number..................Expiry Date...................... signature of the cardholder .................................. Date........................Signature.......................................... _______________________________________________________________________________ _______________________________________________________________________________ _______________________________________________________________________________ International Conference on Molecular Structural Biology Bundeswirtschaftskammer, Vienna, September 17-20, 1995 Hotel Accomodation, All-Inclusive Flight Arrangements, Social Program for accompanying persons REGISTRATION FORM Please return a copy of the completed form to: MONDIAL CONGRESS Faulmanngasse 4 A-1040 Vienna Tel: (43) 1 588 04 -113 or -115 Fax: (43) 1 586 91 85 Mr.... Mrs.... Acad.Degree:..........First Name:................Family Name:................ Dept./Inst.:.......................Street:................................... Postal Code:..........City:......................Country:.................... Phone:.............................Fax:...................................... Hotel Accomodation ...single room ...double room deposit ...Category***** ATS 2170 2530 ATS 2500 ...Category**** ATS 1360 to 1600 1980 to 2110 ATS 2000 ...Category*** ATS 690 to 1050 890 to 1650 ATS 1000 ...Summerhotels*** ATS 550 to 600 760 to 800 ATS 500 ...Summerhotels** ATS 260 400 ATS 500 Date of arrival:.......... Date of departure:.......... Hotel deposit: ATS______ To secure your hotel reservation we kindly ask you to provide us with the deposit listed above. Mondial Congress will be glad to make individual offers for any other hotel upon request. To enable us to book a room for you, please return the attached form and remit the requested deposit by August 4, 1995. You will receive written conformation of your reservation. Shoud the desired hotel category no longer be available, we will do our utmost to offer similar accommodation. This may, however, mean a change of price category. ... Please send me information about the all-inclusive-flight-arrangements (see program) and idicate date of arrival, date of departure and place of departure. Terms of Hotel Cancellation: In case of cancellation or change in bookings, a minimum fee of ATS 500 /person will be charged. For cancellation after August 4, 1995, the hotel deposit will be forfeited. Cancellations or changes must be made in written form. PROGRAM FOR ACCOMPANYING PERSONS No. of Persons ... 'Historical Vienna', Monday, Sept. 18, 1995 ______________ ATS 320 ... 'Art Noveau in Vienna', Monday, Sept. 18, 1995 ______________ ATS 340 ... 'Excursion to the Danube Valley Wachau', Tuesday, Sept. 19, 1995 ______________ ATS 690 ... 'City Walk with a visit to a Viennese Coffee House', Wednesday, Sept. 20, 1995 ______________ ATS 430 EVENING PROGRAM ... Farewell Evening at an Austrian wine tavern "Heurigen", Wednesday, Sept. 20, 1995 ______________ ATS 590 GRAND TOTAL ATS_________ TERMS OF PAYMENT ... remittance to our bank account no. 107-109-499 with the Bank Austria, Wiedner Hauptstrasse, A-1040 Vienna, bank code 12000 "free of charge with the beneficiary" ... send a Eurocheque to MONDIAL CONGRESS, Faulmanngasse 4, 1040 Vienna, together with a copy of the registration form ... Credit Card (please make sure that this form is signed by the credit card- holder) ... Diners Club ... American Express ... Euro/Mastercard ...Visa Credit Card No.:...............................Expiry Date:.............. Signature of the cardholder:.............................................. Date:____________________________ Signature:___________________________________ _______________________________________________________________________________ ALL INCLUSIVE FLIGHT ARRANGEMENTS MONDIAL CONGRESS, in cooperation with the Austrian Airlines and other IATA-carriers is in a position to offer moderately priced all-inclusive flight arrangements to the participants of this congress. These arrangements include the following services: Roundtrip, economy class on regular flights, 20kg (44lbs) luggage allowance, 5 nights, accomodation at **** or *** category hotels in double rooms with private bath and shower, breakfast, service charges, local taxes as well as the transfer airport - hotel. SOCIAL PROGRAM FOR ACCOMPANYING PERSONS September 18, 1995 ------------------ 'Historical Vienna' This tour will show you some of Vienna's most impressive sights including the Ringstrasse with the State Opera House, the National Museums, the Parliament, the City Hall, the Burgtheater, the University and the Votiv Church. Highlight of the tour is a visit to the imperial apartments of Schoenbrunn Palace, the former residence of the Austrian Emperors. On your way back you will pass Belvedere Palace. Duration: 9:00-12:00 Price per person, incl. bus, entrance fees and guide: ATS 320 September 18, 1995 ------------------ 'Art Noveau in Vienna' On this tour you will become acquainted with some of the most important and wellknown buildings of the "Art Noveau" era in Vienna, as well as with some of its great architecs, e.g. Otto Wagner, Josef Hoffmann or Adolf Loos. Highlight of this tour is a visit to the church "Kirche am Steinhof", which has been excellently restored only recently. Duration: 14:00-17:00 Price per person, incl. bus, entrance fees and guide: ATS 340 September 19, 1995 ------------------ 'Excursion to the Danube Valley Wachau' This trip takes you directly to Melk where you will visit the Benedictine Abbey with its famous library and ceiling-frescos. Lunch will be served in the "Stiftstaverne". After lunch we board the steamer going down stream to Duernstein. We round up our excursion with a city stroll through this pittoresque little town with its romantic narrow streets and the well known castle where King Richard Lionhearted was imprisoned. Duration: approx. 8h Price per person, incl. bus, guide, lunch and admission fees ATS 690 September 20, 1995 ------------------ 'City walk with a visit to a Viennese Coffee House' Walking tour through the Hofburg, the imperial winter residence of the Habsburg, and the Treasury, where the insignia of the Holy Roman Empire and the crown of the Austrian Empire illustrate European history. After the visit of the magnificent hall of the National Library, the tour will finish in a typical Austrian coffee shop, where you will taste a "Sachertorte" with coffee or tea. Duration: 14:00-17:00 Price per person, incl. guide, entrance fees, coffee and cake: ATS 430 September 20, 1995 ------------------ 'Farewell evening at an Austrian wine tavern - "Heurigen"' Departure 19:30 from the conference venue. The bus will take you to a wine tavern situated in one of the best known wine growing villages of Vienna. There you will spend There is a opportunity to sumbit late-braking abstracts for presentation at the >> INTERNATIONAL CONFERENCE ON MOLECULAR STRUCTURAL BIOLOGY << Vienna (Austria), September 17-20, 1995 ================================================================================ ================= the DEADLINE is the 7th of AUGUST, 1995.====================== ================================================================================ For details see below: -------------------------------------------------------------------------------- >> INTERNATIONAL CONFERENCE ON MOLECULAR STRUCTURAL BIOLOGY << Organized by the Austrian Chemical Society Vienna (Austria), September 17-20, 1995 TOPICS: 1) The Impact of Molecular Biology on Structural Biology 2) Biomolecular Structure Determination a) X-Ray Diffraction b) NMR Spectroscopy 3) Dynamics and Function of Biomolecules 4) Computational Methods 5) Protein Engineering and Design VIENNA: The organisers would like to invite you to a new conference in the rapidely expanding field of molecular structural biology. The conference will take place in Vienna, home of waltz, the capital of a fomrer great empire, one of the three centres of the United Nations and a leading host city in the world for important meetings and conferences. Scientific topics to be presented at this conference will highlight the state-of-the-art in biomolecular structure determination in its context of molecular biology, medicine and pharmacology. Scientists from both basic and industrial research are highly welcome to present their results to an interested and enthusiastic audience. In addition to the conference itself, you could take time to explore the wonderful city of Vienna, perhaps wandering through the elegant parks in their autumnal colours before soaking up the rich atmosphere of the renowned Viennese cafes. A reception with the mayor in the historic Rathaus at the end of the first day of presentations will provide a taste of the rich Viennese culture waiting to be discovered. ORGANISING COMMITTEE: A.Kungl, P.Andrews, B.Habermann, H.Schreiber, A.Schrenk SCIENTIFIC COMMITTEE: Chairman: P.Schuster M.Breitenbach, C.Kratky, P.Andrews, H.Schreiber A.Kungl IN COOPERATION WITH Austrian Academy of Sciences Federal Ministry of Economy Federal Ministry of Science and Research SPONSORED BY Wirtschaftskammer Oesterreich SCIENTIFIC PROGRAMME: The scientific programme is designed to cover a broad range of disciplines in molecular structural biology. Topics will be presented in the form of plenary lectures, posters, discussions and exhibitions. Sunday, (17.9) ----------------------------------------------------------------------------- Registration Welcome Cocktail Honory Lecture by H.Neurath Monday, (18.9) ----------------------------------------------------------------------------- 1. The impact of Molecular Biology on Structural Biology C.Cantor (USA) "Application of Streptavidin and Genetically Engineered Variants" T.Blundell (GB) "Protein Super Families: Genome Analyses and Drug Discovery" 2. Biomolecular Structure Determination a) X-Ray Crystallography R.Huber (D) "Proteolytic Enzymes and Their Natural Inhibitors - New Functions and New Structures and No End in Sight" M.Walkinshaw (CH) "Structures and Biological Implications of Series of Cyclophilin-Cyclosporin Crystal Complexes" I.Schlichting (D) "Ligand Binding in Heme Proteins Studied by Kinetic X-Ray Crystallography" Tuesday (19.9.) ----------------------------------------------------------------------------- b) NMR Spectroscopy O.Jardetzky (USA) "Flexibility and Function in a DNA Binding Protein - the Trp-Repressor of E.Coli" P.Roesch (D) "The Structure of Iron-Sulfur Proteins in Solution" C.Dobson (GB) "The Structural Basis of Protein Folding" 3. Dynamics and Function of Biomolecules J.Lakowicz (USA) "Fluorescence Spectroscopic Studies of the Structure and Dynamics of Biomolecules" W.Kuehlbrandt (D) "Structure Determination of Membrane Proteins by High-Resolution Electron Microscopy" H.Frauenfelder (USA) "Dynamics and Function of Biomolecules" Wednesday (20.9.) ----------------------------------------------------------------------------- 4. Computational Methods M.Karplus (USA) "Simulations of Protein Folding From the Native to the Denatured State and Back Again" J.Skolnick (USA) "A Hierarchial Approach to the Prediction of Protein Structures" H.-J.Boehm (D) "New Computational Tools for Structure Based Drug Design" 5. Protein Engineering and Design A.Fersht (GB) "Pathways of Protein Folding" L.Presta (USA) "Protein Engineering of Immunoglobulins and Immunoglobulin-Like Domains" M.Mutter (CH) "De Novo Protein Design" CALL FOR POSTERS ----------------- All those intending to participate in the conference are invited to submit posters presenting original work. Abstracts prepared according to the instructions given below should arrive NOT LATER THAN AUGUST 7th, 1995. The authors will be informed about the provisional acceptance in July, 1995. The presentation of the posters will be finally approved and thus the contribution included in the Book of Abstracts when at least one of the registers before August 1st, 1995. Contributors of outstanding posters will be chosen by the scientific committee to give a 20 minute lecture. Abstracts: Camera-ready single-page abstacts of posters should start with the title of the contribution followed by the author(s) name(s), affiliation(s) and address(es). Abstracts must be typed within an area 16cm wide and 24cm long, centred on a sheet of (about) A4 size (210mm x 297mm). The chosen topic must be assigned by stating its title (see list of topics) in the top right margin. The abstracts should be sent unfolded to the conference secreteriat. Proceedings: Invited lectures presented at the conference will be published in book form. All authors will receive in due course instructions for preparing the manuscript (for type-setting, not camera-ready). The deadline for submitting manuscripts is June 30, 1995. Posters: The size of the posters board is 0.9m (width) x 1.5m (height). All posters will be numbered according to their order in the book abstracts. Thumbtacks, tape and markers will be available in the poster area. Head of Scientific Committee: P.Schuster GENERAL INFORMATION: -------------------- EXHIBITON An exhibition of instruments, accessories, software, literature and other items is planned. Companies interested in displaying their products are kindly requested to contact the conference secretariat. INDUSTRIAL SATELLITE MEETING A half day industrial symposium is planned. Those companies who are interested in presenting up-to-date results obtained on their products in the form of 20 minute lectures are kindly requested to contact the conference secretariat. SOCIAL EVENTS The conference will be opened by the welcome cocktail and buffet in the rooms of the Austrian Bundeswirtschaftskammer on Sunday evening, September 17. On Monday, September 18, the mayor of Vienna will invite the conference participants to the historical Viennese Rathaus. A piano concert will take place on Tuesday, September 19, in the hall of the Austrian Gewerbeverein. A farewell evening at a typical Austrian wine tavern is planned for the evening of Wednesday, September 20. Activities for accompanying persons will include cultural events and visits to places of general interest. REGISTRATION To enjoy the early rates, please complete the enclosed Registration Form and send it to the conference secreteriat before August 1, 1995. Registration Fees: before August 1 after August 1 Regular Participant 4000 ATS 4500 ATS GOeCH Member 3500 ATS 4000 ATS Student 2000 ATS 2300 ATS Accompanying Persons 500 ATS 700 ATS The registration desk will be open from 2 p.m. on Sunday, September 17. The registration fee includes the Book of Abstracts, the welcome cocktail and buffet, the invitation to the Viennese Rathaus, a ticket for the piano concert, coffee breaks and tram tickets. ! Meals and accomodation are not included in the registration fee! Tickets for lunch will be available for 190ATS/meal/person (see Registration Form). For accomodation please fill in and return the enclosed flyer of the travel agency MONDIAL CONGRESS to the address given. Remittance of Fees: Remittance must be made in Austrian Schillings (ATS) payalbe to the Gesellschaft Oesterreichischer Chemiker, Abeitsgruppe Biophysikalische Chemie, account number 0043-19265/o4 at Bank Creditanstalt, banking code 11000. Payment is also accepted by sending (Euro-)Cheque to the conference secreteriat or by filling in the credit card form (Visa, Euro/Mastercard, Dinersclub and American Express are accepted). All charges due to bank transfer have to be paid by the sender. The sender's name and address have to be marked clearly on every remittance. Canellation: Applications may be cancelled up to August 15, in which case 85% refund of fees already paid will be made. It will not be possible to offer any refunds if cancellations are received after that date. Key dates: --------- June 30 deadline for submitting poster abstracts and manuscripts August 1 deadline for early registration fee August 15 deadline for cancellation Location: --------- Wirtschaftskammer Oesterreich (Julius Raab Saal) Wiedner Hauptstrasse 63 A-1040 Vienna Conference Secretariat: ---------------------- Gesellschaft Oesterreichischer Chemiker AG Biophysikalische Chemie Dr. Andreas Kungl Nibelungengasse 11 A-1010 Wien, Austria Tel.: (43) 1 587249 FAX.: (43) 1 587966 e-mail: msb95@helix.mdy.univie.ac.at _______________________________________________________________________________ REGISTRATION FORM: Surname........................First Name .....................Title........... Institution/Company............................................................ Departement.................................................................... Street....................................Town................................. Country...................................Post Code............................ Tel.Nr....................................Fax Nr............................... e-mail......................................................................... I am a(n) ...Abstract Submitter ...Invited Speaker ...Regular Participant ...GOECH-Member ...Student Title of Poster/Lecture........................................................ ............................................................................... ...I will require ...Mo ...Tue ...Wed lunch tickets (190 ATS/meal/person) (Payment should accompany registration) Method of Payment: ...Bank Transer to Bank Creditanstalt account number 0043-19265/04, bank code 11000 ...(Euro-)Cheque payalbe to Gesellschaft Oesterreichischer Chemiker, 'International Conference on Molecular Structural Biology' ...Credit Card ...Visa ...Euro/Master ...American Express ...Diners Club Card Number..................Expiry Date...................... signature of the cardholder .................................. Date........................Signature.......................................... _______________________________________________________________________________ _______________________________________________________________________________ _______________________________________________________________________________ International Conference on Molecular Structural Biology Bundeswirtschaftskammer, Vienna, September 17-20, 1995 Hotel Accomodation, All-Inclusive Flight Arrangements, Social Program for accompanying persons REGISTRATION FORM Please return a copy of the completed form to: MONDIAL CONGRESS Faulmanngasse 4 A-1040 Vienna Tel: (43) 1 588 04 -113 or -115 Fax: (43) 1 586 91 85 Mr.... Mrs.... Acad.Degree:..........First Name:................Family Name:................ Dept./Inst.:.......................Street:................................... Postal Code:..........City:......................Country:.................... Phone:.............................Fax:...................................... Hotel Accomodation ...single room ...double room deposit ...Category***** ATS 2170 2530 ATS 2500 ...Category**** ATS 1360 to 1600 1980 to 2110 ATS 2000 ...Category*** ATS 690 to 1050 890 to 1650 ATS 1000 ...Summerhotels*** ATS 550 to 600 760 to 800 ATS 500 ...Summerhotels** ATS 260 400 ATS 500 Date of arrival:.......... Date of departure:.......... Hotel deposit: ATS______ To secure your hotel reservation we kindly ask you to provide us with the deposit listed above. Mondial Congress will be glad to make individual offers for any other hotel upon request. To enable us to book a room for you, please return the attached form and remit the requested deposit by August 4, 1995. You will receive written conformation of your reservation. Shoud the desired hotel category no longer be available, we will do our utmost to offer similar accommodation. This may, however, mean a change of price category. ... Please send me information about the all-inclusive-flight-arrangements (see program) and idicate date of arrival, date of departure and place of departure. Terms of Hotel Cancellation: In case of cancellation or change in bookings, a minimum fee of ATS 500 /person will be charged. For cancellation after August 4, 1995, the hotel deposit will be forfeited. Cancellations or changes must be made in written form. PROGRAM FOR ACCOMPANYING PERSONS No. of Persons ... 'Historical Vienna', Monday, Sept. 18, 1995 ______________ ATS 320 ... 'Art Noveau in Vienna', Monday, Sept. 18, 1995 ______________ ATS 340 ... 'Excursion to the Danube Valley Wachau', Tuesday, Sept. 19, 1995 ______________ ATS 690 ... 'City Walk with a visit to a Viennese Coffee House', Wednesday, Sept. 20, 1995 ______________ ATS 430 EVENING PROGRAM ... Farewell Evening at an Austrian wine tavern "Heurigen", Wednesday, Sept. 20, 1995 ______________ ATS 590 GRAND TOTAL ATS_________ TERMS OF PAYMENT ... remittance to our bank account no. 107-109-499 with the Bank Austria, Wiedner Hauptstrasse, A-1040 Vienna, bank code 12000 "free of charge with the beneficiary" ... send a Eurocheque to MONDIAL CONGRESS, Faulmanngasse 4, 1040 Vienna, together with a copy of the registration form ... Credit Card (please make sure that this form is signed by the credit card- holder) ... Diners Club ... American Express ... Euro/Mastercard ...Visa Credit Card No.:...............................Expiry Date:.............. Signature of the cardholder:.............................................. Date:____________________________ Signature:___________________________________ _______________________________________________________________________________ ALL INCLUSIVE FLIGHT ARRANGEMENTS MONDIAL CONGRESS, in cooperation with the Austrian Airlines and other IATA-carriers is in a position to offer moderately priced all-inclusive flight arrangements to the participants of this congress. These arrangements include the following services: Roundtrip, economy class on regular flights, 20kg (44lbs) luggage allowance, 5 nights, accomodation at **** or *** category hotels in double rooms with private bath and shower, breakfast, service charges, local taxes as well as the transfer airport - hotel. SOCIAL PROGRAM FOR ACCOMPANYING PERSONS September 18, 1995 ------------------ 'Historical Vienna' This tour will show you some of Vienna's most impressive sights including the Ringstrasse with the State Opera House, the National Museums, the Parliament, the City Hall, the Burgtheater, the University and the Votiv Church. Highlight of the tour is a visit to the imperial apartments of Schoenbrunn Palace, the former residence of the Austrian Emperors. On your way back you will pass Belvedere Palace. Duration: 9:00-12:00 Price per person, incl. bus, entrance fees and guide: ATS 320 September 18, 1995 ------------------ 'Art Noveau in Vienna' On this tour you will become acquainted with some of the most important and wellknown buildings of the "Art Noveau" era in Vienna, as well as with some of its great architecs, e.g. Otto Wagner, Josef Hoffmann or Adolf Loos. Highlight of this tour is a visit to the church "Kirche am Steinhof", which has been excellently restored only recently. Duration: 14:00-17:00 Price per person, incl. bus, entrance fees and guide: ATS 340 September 19, 1995 ------------------ 'Excursion to the Danube Valley Wachau' This trip takes you directly to Melk where you will visit the Benedictine Abbey with its famous library and ceiling-frescos. Lunch will be served in the "Stiftstaverne". After lunch we board the steamer going down stream to Duernstein. We round up our excursion with a city stroll through this pittoresque little town with its romantic narrow streets and the well known castle where King Richard Lionhearted was imprisoned. Duration: approx. 8h Price per person, incl. bus, guide, lunch and admission fees ATS 690 September 20, 1995 ------------------ 'City walk with a visit to a Viennese Coffee House' Walking tour through the Hofburg, the imperial winter residence of the Habsburg, and the Treasury, where the insignia of the Holy Roman Empire and the crown of the Austrian Empire illustrate European history. After the visit of the magnificent hall of the National Library, the tour will finish in a typical Austrian coffee shop, where you will taste a "Sachertorte" with coffee or tea. Duration: 14:00-17:00 Price per person, incl. guide, entrance fees, coffee and cake: ATS 430 September 20, 1995 ------------------ 'Farewell evening at an Austrian wine tavern - "Heurigen"' Departure 19:30 from the conference venue. The bus will take you to a wine tavern situated in one of the best known wine growing villages of Vienna. There you will spend an enjoyable evening at the end of this conference with a lavish dinner, wine and typical Viennese music. Duration: 19:30-23:00 Price per person, incl. dinner, 1/2l of wine, mineral water, musical entertainement and bus ATS 590 MONDIAL CONGRESS reserves the right to cancel the tours before the beginning of the congress, should the minimum number of 20 participants not be reached. All tours are subject to change! MONDIAL also offers tickets to concerts and plays in Vienna. If you are interested in some more information please do not hesitate to contact us. an enjoyable evening at the end of this conference with a lavish dinner, wine and typical Viennese music. Duration: 19:30-23:00 Price per person, incl. dinner, 1/2l of wine, mineral water, musical entertainement and bus ATS 590 MONDIAL CONGRESS reserves the right to cancel the tours before the beginning of the congress, should the minimum number of 20 participants not be reached. All tours are subject to change! MONDIAL also offers tickets to concerts and plays in Vienna. If you are interested in some more information please do not hesitate to contact us. -- +-----------------------------------------------------------------------------+ | | | Hellfried Schreiber, Ph.D. | | | +---------------------------------------+-------------------------------------+ | | | | Institute for Theoretical Chemistry | | | Theoretical Biochemistry Group | Mail: hs@helix.mdy.univie.ac.at | | Waehrigerstrasse 17 | Voice: +43 1 40480 - 618 | | A-1090 Wien, Austria, Europe | FAX: +43 1 4028525 | | | | +-----------------------------------------------------------------------------+ From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!daresbury!trane.uninett.no!nntp.uio.no!nac.no!Norway.EU.net!EU.net!howland.reston.ans.net!lamarck.sura.net!darwin.sura.net!nih-csl!loglady.ninds.nih.gov!johnk From: johnk@spasm.niddk.nih.gov (John Kuszewski) Subject: Re: SCIENTIST APPEAL "DON'T LIFT THE TEST BAN" Message-ID: <1995Jul10.171052.18259@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: <3tqml8$ju8@mserv1.dl.ac.uk> Distribution: bionet Date: Mon, 10 Jul 1995 17:10:52 GMT Lines: 24 In article <3tqml8$ju8@mserv1.dl.ac.uk>, JANOT@ill.fr writes: |> Dear Colleague |> About 150 French scientists have signed a petition similar to |> the following in the last few days. We are now sending out this version to Really now. This is both silly and inappropriate for bionet.molbio.proteins. Take it to soc.culture.war.* -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || John Kuszewski || |/ /| || johnk@spasm.niddk.nih.gov || / /|| || \/ / / || \/ that's MISTER protein G to you! |/__/| | /_________| "Biophysics has driven me to an attitude of apocalyptic doom" --Frank Delaglio From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!gatech!EU.net!news.sprintlink.net!dish.news.pipex.net!pipex!sunsite.doc.ic.ac.uk!daresbury!not-for-mail From: JANOT@ill.fr Newsgroups: bionet.molbio.proteins Subject: SCIENTIST APPEAL "DON'T LIFT THE TEST BAN" Date: 10 Jul 1995 08:59:36 +0100 Lines: 96 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3tqml8$ju8@mserv1.dl.ac.uk> Reply-To: JANOT@ill.fr X-Listprocessor-Version: 6.0c -- ListProcessor by Anastasios Kotsikonas Originator: seminars@expga.esrf.fr X-Comment: Interested in seminars at ESRF, Grenoble, France. Errors-To: bertini@esrf.fr Original-To: Multiple recipients of list From: IN%"BERNAS@FRCPN11.IN2P3.FR" 4-JUL-1995 19:48:24.26 To: IN%"behar@if1.if.ufrgs.br" CC: IN%"aguidao@abc.impa.br" Subj: Forwarded Mail Return-path: Received: from diams.ill.fr (DIAMS) by ill.fr (PMDF V4.3-7 #6327) id <01HSHFV7S8RK8Y572X@ill.fr>; Tue, 4 Jul 1995 19:48:18 +1 Received: from out.esrf.fr ([192.168.100.99]) by diams.ill.fr (8.6.9/24Jun94) with ESMTP id TAA04718 for ; Tue, 4 Jul 1995 19:44:39 +0100 Received: (from uucp@localhost) by out.esrf.fr (8.6.10/8.6.10) id TAA01431 for ; Tue, 4 Jul 1995 19:46:48 +0100 Received: from frcpn11.in2p3.fr(134.158.69.66) by firewall via smap (V1.3) id tmp001429; Tue Jul 4 19:46:31 1995 Received: from FRCPN11.IN2P3.FR by frcpn11.in2p3.fr (IBM VM SMTP V2R2) with BSMTP id 0577; Tue, 04 Jul 95 19:45:02 SET Received: from FRCPN11.IN2P3.FR (NJE origin BERNAS@FRCPN11) by FRCPN11.IN2P3.FR (LMail V1.2a/1.8a) with BSMTP id 3662; Tue, 4 Jul 1995 19:45:02 +0200 Date: Tue, 04 Jul 1995 19:44:34 +0000 (SET) From: BERNAS@FRCPN11.IN2P3.FR Subject: Forwarded Mail To: behar@if1.if.ufrgs.br Cc: aguidao@abc.impa.br Message-id: <199507041846.TAA01431@out.esrf.fr> Organization: In2p3 MIME-version: 1.0 Content-type: TEXT/PLAIN; CHARSET=US-ASCII Content-transfer-encoding: 7BIT ----------------------------Original message---------------------------- Date: Mon, 03 Jul 95 12:53:56 SET From: BERNAS@FRCPN11.IN2P3.FR Organization: In2p3 To: ALL COLLEAGUES Message-Id: <950703.135452.SET.BERNAS@FRCPN11.IN2P3.FR> Dear Colleague About 150 French scientists have signed a petition similar to the following in the last few days. We are now sending out this version to our colleagues all over the world. The undersigned will be pleased to record your reaction. Thank you. ----------------------------------------------------------------------------- SCIENTISTS APPEAL "DON'T LIFT THE TEST BAN" We are scientists and citizens. We wish to state our firm opposition to the lifting of the nuclear test ban, by the French government or any other. Several countries already have large nuclear arsenals, and we all know that the number of existing warheads is large enough to destroy the Earth several times over. The problem today is not to maintain or improve them, but get rid of them. today, we live in a different world from that in which the arms race led to the nuclear stalemate. The world we now live absolutely requires that all of mankind face the need to put an end - a final end - to the pileup of massively destructive arms. This is now politically and economically necessary, and indeed nuclear stockpiles are being progressively reduced all over the world in spite of many difficulties and hesitations. It is technically possible to enhance the rate at which these arms are dismantled. The timely goal should now be their total elimination. The decision of President Chirac to resume atomic weapons tests in 1995 and 1996 is thus dramatically retrograde. We are concerned that the present "limit- ed" weapons test program envisaged by several nuclear powers and whose implem- entation has just been decided by France could well be aimed at adjusting some parameters in simulations that are preparing new weapons. The pressure of nuc- lear arms lobbies is mounting in many countries, and many experts have confirm- ed that - with simulations - 4 to 10 tests are enough to design a new weapon. Is France, are other governments, going to turn History's wheel backward by re- suming the arms race ? Mankind deserves a better future. We are aware that the destruction of Hiroshima and Nagasaki, and the const- ant development of new "high-tech" weapons, have done much to spoil the image of scientific progress. A new weapons race not only carries the dangers of phy- sical destruction. Even if no bomb goes off, the fear generated by eapons tech- nology can affect mankind's will to understand our world and to use science as a tool to solve it's huge problems in food production, health and education, to find new ways to protect and develop soils, renew energy resources and reduce pollution. And, of course, the money that goes into sterile bombs is money that does not help to solve any of these problems - just makes them worse. For these reasons, we publicly request that the nuclear test moratorium be maintained. --------------------------------------------------------------------- For information and to join this appeal, contact P. Jaegle or A. Sureau, Laboratoire de Spectroscopie Atomique et Ionique, Universite Paris-Sud, 91405- Orsay, France, EMAIL = pierre.jaegle@lsai.u-psud.fr OR H. Bernas, CSNSM, 91405 Orsay, France, EMAIL = bernas@frcpn11.in2p3.fr From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!rutgers!uwm.edu!news.moneng.mei.com!news.ecn.bgu.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!uunet!in1.uu.net!cis.ohio-state.edu!magnus.acs.ohio-state.edu!mhsieh From: mhsieh@magnus.acs.ohio-state.edu (Ming-Ching Hsieh) Newsgroups: bionet.molbio.proteins Subject: Help! A Clogged HIC Cartridge (Bio-Rad) Date: 11 Jul 1995 03:33:51 GMT Organization: The Ohio State University Lines: 15 Distribution: usa Message-ID: <3tsrev$s6b@charm.magnus.acs.ohio-state.edu> NNTP-Posting-Host: beauty.magnus.acs.ohio-state.edu Hi Dear Netters: Can anyone tell me how to recover a clogged HIC cartridge, which is an Econo-Pac Methyl HIC cartridge made by Bio-Rad? I'm using it to purify a plant apoplastic protein. I broght the infiltration solution to 2.4 M ammonium sulfate and applied it to the cartridge. The proteins were eluted with a linear gradient of ammonium sulfate from 2.4 M to zero. The specific enzyme came out at the end of gradient. In the beginning, the flow rate was about 1 ml/min. However, after several uses, the flow rate was reduced to 1 ml/hr. I did clean the cartridge as suggested by the manufacturer, using 100mM NaOH, 20% glycerol, and deionized water to wash. And I even washed it with 0.5M NaOH and 70% ethanol. They all didn't work. Please tell me what else I can do. Thanks a lot in advance. Ming hsieh.21@ous.edu From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!rutgers!gatech!newsxfer.itd.umich.edu!isclient.merit.edu!cwis-20.wayne.edu!usenet From: Anton Scott Goustin Newsgroups: bionet.molbio.proteins,bionet.software Subject: Re: Program to calculate isoelectric points? Date: 11 Jul 1995 00:59:22 GMT Organization: Wayne State University Lines: 16 Message-ID: <3tsida$qtg@cwis-20.wayne.edu> References: NNTP-Posting-Host: dyn51-49.biosci.wayne.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: quoted-printable X-Mailer: Mozilla 1.1N (Macintosh; I; PPC) X-URL: news:DBIKrp.AAn@midway.uchicago.edu Xref: biosci bionet.molbio.proteins:5020 bionet.software:12656 scb1@ellis.uchicago.edu (Samuel C. Blackman) wrote: >Does anyone know of a program (Mac, DOS, Unix) that will calculate the >isoelectric point of a given peptide? Thanks! > >-- Sam > > >-- >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 There is a program that does so contained within MacVector 4.5 (Kodak Imaging Technologies)...expensive. Perhaps a neighbor has it = there at the University of Chicago. From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!newsserver.jvnc.net!yale.edu!news.ycc.yale.edu!news From: Edward C. Goodwin, Ph.D. Newsgroups: bionet.molbio.proteins Subject: Wanted-Antibodies to Lac Repressor or HSV-VP16 Date: 10 Jul 1995 21:04:06 GMT Organization: Yale University Lines: 14 Distribution: world Message-ID: <3ts4k6$17v@news.ycc.yale.edu> NNTP-Posting-Host: 130.132.88.91 X-Newsreader: Nuntius Version 1.2 X-XXDate: Mon, 10 Jul 1995 21:10:28 GMT I'm trying to use an inducable system based on a Lac Repressor-Herpes VP16 fusion protein. However, I really need to document expression of this protein since functional assays are a pain and need to be reserved for a few stable cell lines. A western would seem to be ideal but I am having an unexpectedly difficult time in tracking down antibodies to Lac I or VP16 (I've checked the usual sources without success). If you know of any such antibodies, or better yet have an excess that you'd be willing to share, please drop me a line. Thanks in advance Ed Edward C. Goodwin Ph.D. ecg@po.cwru.edu Yale University Department of Genetics From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!Germany.EU.net!EU.net!uknet!bhamcs!news.ox.ac.uk!oxpath!casselton From: casselton@molbiol.ox.ac.uk Newsgroups: bionet.molbio.proteins Subject: homeodomain proteins-postdoc Date: 10 Jul 95 21:17:05 GMT Organization: Oxford University Molecular Biology Data Centre Lines: 19 Message-ID: <1995Jul10.211705@ania.path.ox.ac.uk> NNTP-Posting-Host: ania.path.ox.ac.uk POSTDOCTORAL RESEARCH ASSISTANT - FUNGAL MOLECULAR GENETICS Applications are invited for a postdoctoral position to study the interactions of homeodomain proteins that regulate sexual development and determine self-nonself recognition in the mushroom Coprinus cinereus. The post is funded by BBSRC for two years and is available immediately. Salary will be on the RS 1A scale. The project is jointly supervised by Dr Lorna Casselton in the Department of Plant Sciences and Dr Jane Mellor in the Department of Biochemistry. Applications, including a detailed curriculum vitae and names of two referees, should be sent to Administrator, Department of Plant Sciences, South Parks Road, Oxford OX1 3RB. Closing Date 18 August 1995. Enquiries: Lorna Casselton Tel. 01865 275 109; FAX 01865 275 074; e mail Casselton@molbiol.ox.ac.uk From owner-proteins@net.bio.net Sun Jul 09 23:00:00 1995 Newsgroups: bionet.molbio.proteins,bionet.software Path: biosci!rutgers!gatech!news.uoregon.edu!vixen.cso.uiuc.edu!uchinews!ellis!scb1 From: scb1@ellis.uchicago.edu (Samuel C. Blackman) Subject: Program to calculate isoelectric points? 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: Mon, 10 Jul 1995 18:59:48 GMT Lines: 11 Xref: biosci bionet.molbio.proteins:5015 bionet.software:12647 Does anyone know of a program (Mac, DOS, Unix) that will calculate the isoelectric point of a given peptide? Thanks! -- Sam -- 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 Sun Jul 09 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!swrinde!dish.news.pipex.net!pipex!sunsite.doc.ic.ac.uk!qmw!orac.sunderland.ac.uk!usenet From: len.bell@Sunderland.AC.UK (Anonymous) Newsgroups: bionet.molbio.proteins Subject: metal peptide coordination Date: 10 Jul 1995 07:46:11 GMT Organization: University of Sunderland Lines: 14 Message-ID: <3tqls3$p79@orac.sunderland.ac.uk> Reply-To: hs0lbe@orac.sund.ac.uk NNTP-Posting-Host: glaxo3.sunderland.ac.uk X-Newsreader: WinVN 0.91.4 i am interested in the coordination of metal ions, zinc and copper, by peptides. and would be interested to hear about good reveiw articles on the subject as well as general information. My particular interest would be for information concerning the structural alterations that occur upon coordination. helix-coil- beta sheet and the like, and especially how these changes could be investigated. i am aware of the fact that predominately histidine and glutamic acid are the favoured amino acid residue ligands, but information about the coordination geometry and structure would be greatly appreciated. thanks len. From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!news.uoregon.edu!news.bc.net!info.ucla.edu!nnrp.info.ucla.edu!usenet From: arne@hodgkin.mbi.ucla.edu (Arne Elofsson) (Arne Elofsson) Newsgroups: bionet.molbio.proteins Subject: Re: Is the GD Rose paper out? Date: 11 Jul 1995 23:36:19 -0700 Organization: University of California, Los Angeles Lines: 63 Message-ID: <6g7n5o33bw.fsf@hodgkin.mbi.ucla.edu> References: <3tucm1$gg6@agate.berkeley.edu> <3tug4i$7jg@nexus.uiowa.edu> NNTP-Posting-Host: hodgkin.mbi.ucla.edu In-reply-to: kpmurphy@blue.weeg.uiowa.edu's message of 11 Jul 1995 18:32:50 GMT X-Newsreader: (ding) Gnus v0.91 In article <3tug4i$7jg@nexus.uiowa.edu> kpmurphy@blue.weeg.uiowa.edu (murphy kenneth p) writes: > > In article <3tucm1$gg6@agate.berkeley.edu>, > Louis Hom wrote: > >I was just wondering if that paper by Rose et al. regarding The Problem had > >come out yet. I seem to recall someone predicting its publication sometime > >last month or this month. Medline showed nothing. > >-- > >______________________________________________________________________________ > >Lou Hom >K'93 I hear the American Enterprise Institute > >lhom@ocf.berkeley.edu says it's true. > >http://www.ocf.berkeley.edu/~lhom/ > > Yes, it's in the latest issue of Proteins as the lead article. > so what did you gues think. Is in time to try to join the AVP beachvolleyboll teams, or should I continue my science carrier ? Well I'm starting my ow lab next month so I guess that is my answer. However if you calculate the accuracy of secondarystructure prediction in the 5 proteins reported is 92 % (and if you do not include apo-myoglobin it is it is 94 %). Which is more or less as good as you get if you compare for instance between procheck and dssp for the same protein, and better than between highly related structures. But this is for only four (different) proteins. On the other side the packing of secondary structure elements is not that impressive. Looks pretty good for the fragments in fig 3 (1fb) and for 2 out of three 50 aa fragments of b562. but pcy, mbo and eglin does not look that good (when you take into aspect that you have nearly perfect secondary structure prediction). MBO is not a very good test as apomyoglobin not is stable in water. It is interesting that such a simple method seems to work that well. Unfortunately this is one of many studies that would be extremely much better if they had done some more rigourous testing, from what I read in the John Hopkins journal, the run takes over night or so, which makes me wonder why they do not run this on 50 different proteins. I always get very very suspicious abot this method. How do I know that they did not try it on 50 proteins and it only worked in these 5 ? Probably not but they might have unintentionally have optimised things for this test set. Any study that do not contain a jack-knife test is much less impressive to me. But atleast it gives me a few new ideas and I guess that makes it a good paper. It is also described (more or less) good enough so that anyone can implement the algorithm and try it themself. 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 Mon Jul 10 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!gatech!news.sprintlink.net!EU.net!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: cpnintt@aol.com (CPN INTt) Newsgroups: bionet.molbio.proteins Subject: Need Purified Protein - Neutral or Alkaline Cellulase Date: 12 Jul 1995 00:20:31 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 10 Sender: root@newsbf02.news.aol.com Message-ID: <3tviif$35q@newsbf02.news.aol.com> Reply-To: cpnintt@aol.com (CPN INTt) NNTP-Posting-Host: newsbf02.mail.aol.com We are looking for purified betaglucanases, purified endoglucanases and cellobiohydralases which are active at pH 7.0 and/or pH 10.0 at temperatures between 40 - 60 C If you have any interesting candidates please send us info to fax # 1-407-743-8343 or our direct e.mail. rgds MAE From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Newsgroups: bionet.molbio.proteins Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!uchinews!drugs-mac16.bsd.uchicago.edu!user From: r-litt@uchicago.edu (Bob Litt) Subject: Re: Triton-X 100 effect on protein concentration determination X-Nntp-Posting-Host: drugs-mac16.bsd.uchicago.edu Message-ID: Sender: news@midway.uchicago.edu (News Administrator) Organization: Univ of Chicago References: <3thfrq$6t3@sifon.cc.mcgill.ca> Date: Wed, 12 Jul 1995 02:56:07 GMT Lines: 22 In article , spang@vms.biochem.mpg.de (Anne Spang) wrote: > Hi! > Triton X-100 messes up your Bradford assay! In the presence of detergent > in our sample, we usually precipitate the sample with TCA in a dotblot > apparatus on nitrocellulose, then stain with amido black. After elution > teh samples can be read in an ELISA reader or a normal photometer. > > Hope this helps > Anne I have heard that the BioRad DC assay is supposed to be compatible with triton X-100. Has anyone tried this, and does it work as advertised? Bob Litt -- r-litt@uchicago.edu Segev Lab Department of Molecular Genetics and Cell Biology University of Chicago From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!rutgers!gatech!news.uoregon.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!EU.net!Austria.EU.net!newsfeed.ACO.net!swidir.switch.ch!scsing.switch.ch!news.belwue.de!News.Uni-Marburg.DE!news.th-darmstadt.de!fauern!lrz-muenchen.de!news From: Wolfgang Schechinger Newsgroups: bionet.molbio.proteins Subject: Re: Triton X-100, TCA, Protein determination Date: 11 Jul 1995 20:44:14 GMT Organization: Leibniz-Rechenzentrum, Muenchen (Germany) Lines: 8 Distribution: world Message-ID: <3tunqu$mcp@sparcserver.lrz-muenchen.de> References: NNTP-Posting-Host: u7k0201.ppp.lrz-muenchen.de Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: spang@vms.biochem.mpg.de Danke ! Wolgang Schechinger Institut für Diabetesforschung Kölner Platz 1 München Fon 089 / 3079 3124 From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!rutgers!gatech!news.uoregon.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!EU.net!Austria.EU.net!newsfeed.ACO.net!swidir.switch.ch!scsing.switch.ch!news.belwue.de!News.Uni-Marburg.DE!news.th-darmstadt.de!fauern!lrz-muenchen.de!news From: Wolfgang Schechinger Newsgroups: bionet.molbio.proteins Subject: Re: Help! A Clogged HIC Cartridge (Bio-Rad) Date: 11 Jul 1995 20:41:46 GMT Organization: Leibniz-Rechenzentrum, Muenchen (Germany) Lines: 8 Distribution: world Message-ID: <3tunma$mcp@sparcserver.lrz-muenchen.de> References: <3tsrev$s6b@charm.magnus.acs.ohio-state.edu> NNTP-Posting-Host: u7k0201.ppp.lrz-muenchen.de Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: mhsieh@magnus.acs.ohio-state.edu Dear Ming, Maybe you will be successful if you reverse the direction of flow while cleaning the column? Good luck! Wolfgang Schechinger Institute for Diabetes Research Munich From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: "Andrew, Tel. +396-91093434" Newsgroups: bionet.molbio.proteins Subject: RE: metal peptide coordination Date: 11 Jul 1995 19:20:45 +0100 Lines: 26 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3tufdt$1uc@mserv1.dl.ac.uk> Original-To: proteins@dl.ac.uk Hi Len, I have to post this here as a reply to your own address bounced. Anyway, zinc fingers might be one possibility to look at - zinc ion coordination is required for their DNA-binding domains to fold correctly. The relevant binding domains are stable as shortish peptides (20-30 aas long) and display a rapid cooperative transition from a random coil to a 2-stranded-beta sheet - alpha-helical structure upon binding to metal ion, which can be followed by circular dichroism and UV-spectroscopy (the latter if cobalt is used instead of zinc). Look for papers by Jeremy Berg (a Medline search should turn him up) and there are also several X-ray and NMR structures of zinc fingers and steroid hormone receptors (which use similar domains) in the PDB database where you can extract geometrical information. All the best, Andrew =============================================================================== Andrew Wallace, IRBM P. Angeletti, Pomezia, Italy. Voice: +39-6-91093434 Fax: +39-6-91093225 Email: wallace@irbm.it Discussion on phage display, combinatorial libraries, etc. - molreps@irbm.it DISCLAIMER: I speak only for myself. =============================================================================== From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!agate!lhom From: lhom@OCF.Berkeley.EDU (Louis Hom) Newsgroups: bionet.molbio.proteins Subject: Is the GD Rose paper out? Date: 11 Jul 1995 17:33:53 GMT Organization: U. C. Berkeley Open Computing Facility Lines: 8 Message-ID: <3tucm1$gg6@agate.berkeley.edu> NNTP-Posting-Host: lightning-ether.berkeley.edu I was just wondering if that paper by Rose et al. regarding The Problem had come out yet. I seem to recall someone predicting its publication sometime last month or this month. Medline showed nothing. -- ______________________________________________________________________________ Lou Hom >K'93 I hear the American Enterprise Institute lhom@ocf.berkeley.edu says it's true. http://www.ocf.berkeley.edu/~lhom/ From owner-proteins@net.bio.net Mon Jul 10 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: A reference for y'all Message-ID: <1995Jul11.164107.14941@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, 11 Jul 1995 16:41:07 GMT Lines: 17 Srinivasan & Rose, Proteins 22:81-99 (1995) -- _____________ | ___/_ | |/ / -- /\ // /-- || || / /|| || || / / || || ||/ / || 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 Jul 10 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: Mohamed SAAD ESRF BP 220 38043 GRENOBLE CEDEX Newsgroups: bionet.molbio.proteins Subject: nuclear tests (fwd) Date: 11 Jul 1995 14:38:39 +0100 Lines: 87 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3ttusv$i13@mserv1.dl.ac.uk> Original-To: proteins@dl.ac.uk ================== Hi folks, here is something important..... ----- Begin Included Message ----- # # # # #### # ###### ## ### ## # # # # # # # # # # # # # # # # # # ##### # # # # # # # # # # # # ###### ##### # ## # # # # # # # # # # # # #### #### ###### ###### # # # # ##### ###### #### ##### #### # # # # # # # # ##### #### # #### # # # # # # # # # # # # # # # ###### #### # #### # 1 SHIMIZU Seishi Physics,University of Tokyo,Japan 2 Yuichi Nishihara Physics,University of Tokyo,Japan 3 Hirohisa TANIGUCHI Physics,University of Tokyo,Japan 4 Takashi Tomoeda Physics,University of Tokyo,Japan 5 Tomoki KOBAYASHI Physics,University of Tokyo,Japan 6 Munehito ARAI Physics,University of Tokyo,Japan 7 Akira Okazaki Physics,University of Tokyo,Japan 8 Atsushi Matsumura Physics, Tohoku University, Japan 9 Kouta Yamamoto Chemistry,Tohoku University,Japan 10 Yasushi UJIOKA Degremont S.A., France 11 Toru Hara Universite de Paris Sud, France 12 Rene Bakker CEA - Sacley, France 13 David Garzella Universite de Paris Sud, France 14 Henk Blok Vrije Universiteit/NIKHEF, Amsterdam 15 Igor Passchier NIKHEF, Amsterdam 16 Ard van Sighem NIKHEF, Amsterdam 17 Johan Noordhoek KOL Leiden 18 C.M.C.M. van Woerkens Kamerlingh Onnes Laboratory, Leiden 19 Annemarie Borst, Vrije Universiteit Amsterdam 20 Gijs Nelemans Universiteit Utrecht 21 Susanne Buiter Universiteit Utrecht 22 Stan Schoofs Universiteit Utrecht 23 Edward Prendergast Universiteit Utrecht 24 Manon Kluytmans Universiteit Utrecht 25 Edmar Weitenberg Utrecht 26 Harry Blom Ruimteonderzoek Utrecht 27 Henk Marquering Seismology, Utrecht 28 Marlies ter Voorde Amsterdam 29 Anco Lankreijer Amsterdam 30 Hans Veldkamp Bilthoven 31 Lyande Eelderink Enschede 32 Christine Pohl Enschede, NL 33 Gottfried Schneiders DLR, Oberpfaffenhofen, Germany 34 Susanne Lehner DLR, Oberpfaffenhofen, Germany 35 Miriam de las Heras, AWI, Bremerhaven, Germany 36 Henning Stahlberg, University Lausanne, Switzerland 37 Oliver Kohle, University Lausanne, Switzerland 38 Angelika Priese, University Hamburg, Germany 39 Klaus-Dieter Liss, European Synchrotron Radiation Facility ESRF, Grenoble,F 40 SAAD Mohamed,Grenoble,F Dear Sirs, This is a chain letter to urge the french government to stop nuclear tests. If you agree with us, please add your name to the list above, and send copies to your freinds. We will add up the lists that had come back to us, and send it to the French Government. If you happen to be the hundredth,two hundredth, three hundredth, and so on, on the list, please send a copy of the mail back to the addresses below, so that we can keep track of this project. If you have any comment please send mails to us. And also, if you are multi-lingual and have friends who may not understand English, please translate this message and add it to the end of the mail. Thank you very much. ******* addresses of the organizers shimizu@femto.phys.s.u-tokyo.ac.jp keshi@uticeaix1.icepp.s.u-tokyo.ac.jp <- please use this adress ******* ----- End Included Message ----- From owner-proteins@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!rutgers!oitnews.harvard.edu!fas-news.harvard.edu!n