From owner-7tms_r@net.bio.net Wed Feb 01 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: Cornelius Krasel Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Helix 7 & C-terminal tails Date: 2 Feb 1995 20:29:29 -0000 Lines: 27 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3grfb9$sbg@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Yoshihisa Inoue (inoue@greencross.co.jp) wrote on 19 Jan 95: [lots deleted ... have still to keep up with the literature :-( ] > And M.Mahmoudian reported "the > complex of human Gs protein with the beta3 adrenergic receptor" > (J.Mol.Graphics,12,22-28 & 34 (1994). Has anybody compared the postulated structure of Gs with the published structures of Gi and Gt? I am always very unsure when I read structure predictions because I have no idea at all how far I can trust them. Maybe the comparison would give an idea of how reliable such a prediction might be. As for the other papers quoted by Dr. Inoue -- I still have to read most of them :-( I would like to see this thread keeping alive since I think interaction of GPCRs with other proteins is one of the most interesting areas in the field :-) (Disclaimer: I'm not working in this area.) --Cornelius. -- /* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */ /* email: krasel@alf.biochem.mpg.de fax: +49 89 8578 3795 */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Wed Feb 01 22:00:00 1995 Path: biosci!PHARMACOP.COM!inglese From: inglese@PHARMACOP.COM (Jim Inglese) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 2 Feb 1995 16:47:09 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Subscribe From owner-7tms_r@net.bio.net Thu Feb 02 22:00:00 1995 Path: biosci!RULGCA.LEIDENUNIV.NL!IJZERMAN From: IJZERMAN@RULGCA.LEIDENUNIV.NL Newsgroups: bionet.molbio.proteins.7tms_r Subject: atomic coordinates of Gi Date: 3 Feb 1995 04:20:19 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HMLZI0S4OY0002JI@rulgca.LeidenUniv.nl> Dear netters, I am looking for the atomic coordinates of the crystal structures of Gi as reported on by Coleman and others. In the PDB these structures are still 'on hold'. Is it possible to retrieve them from somewhere else? Thanks for your help Ad IJzerman From owner-7tms_r@net.bio.net Thu Feb 02 22:00:00 1995 Path: biosci!greencross.co.jp!inoue From: inoue@greencross.co.jp (Yoshihisa Inoue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang-II non-peptide Agonist Date: 3 Feb 1995 21:01:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 59 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502040500.OAA10889@gccss2.greencross.co.jp> NNTP-Posting-Host: net.bio.net Dear Netters, I think you have already read the following interesting paper. AU S.Perlman, H.T.Schambye, R.A.Rivero, W.J.Greenlee, S.A.Hjorth, and T.W.Schwartz TI Non-peptide angiotensin agonist. Functional and molecular interaction with the AT1 receptor. SO J.Biol.Chem.,270(04).1493-1496. The points are: (1) Merck group found non-peptide antagonists L-162313. (2) In the COS-7 cells, the maximal response of IP accumuration was 50%, and in the CHO cells, 3% of the maximal A-II response. (3) L-162313 is a partial agonist in CHO or COS-7 cells, but in vivo it increased blood pressure to the same degree of A-II, but the onset is slow and duration is long. The increase of the blodd pressure is not caused only by the docking of A-II with the receptor, but by the cascade after the dock. (4) The binding sites of A-II, L-162313, and antagonist L-158809 are slightly different. And you have to remember that the competitive and insurmountable antagonists were affected differently by the mutation (by Schambye et al, Br.J.Pharmacol). What mechanism? I have an idea but difficult for me to explain in English. By the way, the following 3 articles are also interesting. AU H.Sasamura, V.J.Dzau, and R.E.Pratt TI Desensitization of Angiotensin Receptor Function SO Kidney International, 46, 1499-1501 (1994). AU L.Hunyady, Y.Tian, K.Sandberg, T.Balla, and K.J.Catt TI Divergeent Conformational Requirements for Angiotensin II Receptor Internalization and Signaling SO Kidney International, 46, 1496-1498 (1994). AU S.Chaki, D.-F.Guo, Y.Yamano, K.Ohyama, M.Tani, M.Mizukoshi, H.Shirai, and T.Inagami TI Role of Carboxylic Tail of the rat Angiotensin II Type 1a Receptor in Agonist-induced Internalization of the Receptor SO Kidney International, 46, 1492-1495 (1994). The first one describes that the PKC is involved in the desensitization, and because the rate of dissapearance of the receptor from the cell surface is considerably slower than the rate of agonist-induced desensitizaion and so on, the process of the sequestration/internalization is different. The second one describes that because the NPxY sequence present in the cytoplasmic tail of EGF or insulin receptors functions as internalization signal, NPxxY in TM7 might be the strong candidate. The third one describes that the C-tail is important for the internali- zation. ____/ ___/ ___/ Yoshihisa INOUE (^_^) the Green Cross Corp. / / / 2-25-1 Shodai-Ohtani,Hirakata,Osaka 573 JAPAN / _ / / / tel: +81-720-56-9328 / / / / fax: +81-720-68-9597 _____/ _____/_____/ E-mail: inoue@greencross.co.jp From owner-7tms_r@net.bio.net Fri Feb 03 22:00:00 1995 Path: biosci!uni-duesseldorf.de!schuessl From: schuessl@uni-duesseldorf.de (Peter Schuessler) Newsgroups: bionet.molbio.proteins.7tms_r Subject: g-proteins Date: 4 Feb 1995 04:53:31 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502041253.EAA10437@net.bio.net> NNTP-Posting-Host: net.bio.net hello, can everyone give me more information about the interaction of a Gs alpha protein and the membrane. Is it right that this interaction is based on hydrophobic portions of the G protein? And is it right that all G alpha proteins assign to the membrane associated protein fraction instead of the membrane-integrated fraction or the cytosolic fraction? I'am interested in this points because I will use a Gs alpha protein antibody to detect the membrane associated proteins and discriminate them from the outher fractions after a separation procedure of whole proteins into these different soluble phases. Thank you very much With Best Regards, Peter Schuessler Peter Schuessler Institut fuer Genetik Heinrich-Heine-Universitaet Duesseldorf D-40225 Duesseldorf Germany Tel: ++49-(0)211-311-2408 e-mail: schuessl@mail.rz.uni-duesseldorf.de From owner-7tms_r@net.bio.net Fri Feb 03 22:00:00 1995 Path: biosci!rutgers!concert!news-server.ncren.net!news.duke.edu!agate!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: g-proteins Date: 4 Feb 1995 14:20:16 GMT Organization: University of Michigan Lines: 48 Message-ID: <3h02f0$3r5@lastactionhero.rs.itd.umich.edu> References: <199502041253.EAA10437@net.bio.net> NNTP-Posting-Host: warbler.med.umich.edu schuessl@uni-duesseldorf.de (Peter Schuessler) wrote: > can everyone give me more information about the interaction of a Gs > alpha protein and the membrane. Is it right that this interaction is based > on hydrophobic portions of the G protein? And is it right that all G alpha > proteins assign to the membrane associated protein fraction instead of the > membrane-integrated fraction or the cytosolic fraction? The relative distribution of G proteins between plasma membrane, intracellular membranes, and cytosol is still rather confusing and is probably different for different G proteins. I recently reviewed some of the literature on this topic in: FASEB J 8:939-946 (1994) Membrane organization in G protein mechanisms. > > I'am interested in this points because I will use a Gs alpha protein > antibody to detect the membrane associated proteins and discriminate them > from the outher fractions after a separation procedure of whole proteins > into these different soluble phases. There have since been some nice papers on the role of palmitoylation of Galpha-s specifically in the interaction of that protein with membranes. If you look at Wedegaertner PB Bourne HR Activation and depalmitoylation of Gs alpha. Cell 77:1063-70, (1994) I think you will find that using Gs as a marker for plasma membrane will not be valid as agonist activation of Gs results in loss of palmitoylation and release from the plasma membrane into a cytosolic fraction. We have also looked at the distribution of labelled alpha and beta-gamma subunits introduced by membrane fusion into NG-108-15 cells and found that while beta-gamma stayed in the plasma membrane the alpha subunit (alpha-o in this case) was also present intracellularly but appeared to be in either a membrane bound or aggregated state as it pelleted on centrifugation. Kwon et al Cell. Signal. 6:663-679 (1994). Lateral mobility of tetramethylrhodamine-labelled G protein a and bg subunits in NG 108-15 cells. Cheers, Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Fri Feb 03 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: Cornelius Krasel Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: g-proteins Date: 4 Feb 1995 14:45:54 -0000 Lines: 30 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3h03v2$dpi@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Peter Schuessler (schuessl@mail.rz.uni-duesseldorf.de) wrote: >can everyone give me more information about the interaction of a Gs >alpha protein and the membrane. Is it right that this interaction is based >on hydrophobic portions of the G protein? And is it right that all G alpha >proteins assign to the membrane associated protein fraction instead of the >membrane-integrated fraction or the cytosolic fraction? I think it is not entirely clear how the interaction between G alpha subunits and membranes takes place. It has been shown that, in the case of Gs, palmitoylation of the alpha subunit is necessary for its membrane targeting (Wedegaertner et al., JBC 268, 25001-25008, 1993). Upon stimulation of Gs-coupled receptors the palmitate can get lost, resulting in trans- location of the Gs alpha subunit into the cytosol (Wedegaertner & Bourne, Cell 77, 1063-1070, 1994). On the other hand, the contrary has been reported, namely that stimulation of Gs with cholera toxin (Degtyarev et al., JBC 268, 23769-23772, 1993) or b2-AR agonists (Mumby et al., PNAS 91, 2800-2804, 1994) _increases_ palmitoylation of the alpha subunit. A review about lipid modifications of trimeric G proteins can be found in JBC 270 (2), 503-506, 1995 (by Wedegaertner, Wilson and Bourne). Hope this is of some help, --Cornelius. -- /* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */ /* email: krasel@alf.biochem.mpg.de fax: +49 89 8578 3795 */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Fri Feb 03 22:00:00 1995 Path: biosci!rutgers!csn!magnus.acs.ohio-state.edu!usenet.ins.cwru.edu!agate!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: atomic coordinates of Gi Date: 4 Feb 1995 14:01:01 GMT Organization: University of Michigan Lines: 18 Message-ID: <3h01at$3r5@lastactionhero.rs.itd.umich.edu> References: <01HMLZI0S4OY0002JI@rulgca.LeidenUniv.nl> NNTP-Posting-Host: warbler.med.umich.edu IJZERMAN@RULGCA.LEIDENUNIV.NL wrote: > I am looking for the atomic coordinates of the crystal structures of Gi as > reported on by Coleman and others. In the PDB these structures are still 'on > hold'. Is it possible to retrieve them from somewhere else? > Thanks for your help In general until things are officially released you need to contact the authors to obtain the coordinates. I was able to get the transducin alpha coordinates from the Sigler group before they were released to the public just by asking. You may need to be persistent. I'd suggest that you contact Sprang or Gilman to ask if they will give them to you. "Legally" they aren't required to until the 1 year hold on the coordinates is over. Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!TITUS.U-STRASBG.FR!phili From: phili@TITUS.U-STRASBG.FR Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 6 Feb 1995 06:00:22 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502061400.AA21938@titus> NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!pipex!lyra.csx.cam.ac.uk!warwick!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (AJ. Collier) Subject: membrane spanning domains Message-ID: Sender: usenet@info.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: zeus.bris.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL2] Date: Mon, 6 Feb 1995 15:39:02 GMT Lines: 7 Does anyone know of a G-protein linked receptor which doesnt possess 7 membrane spanning domains. Thanks Adam Collier :quit From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 6 Feb 1995 17:08:37 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 57 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net > > Dear Dr.Dave Kristofferson, > > I am interested in G-protein Coupled Receptor, so I join 7tms_r Mailing > list, and also read bionet.molbio.proteins.7tms_r. But it is strange. > Suppose that I post some comments to 7tms_r@net.bio.net, and it is > distributed to the "Mailing List" people, but not appeared in the > "Bionet.molbio.proteins.7tms_r" Newsgroup. We tested the system previously and it worked fine. I have just posted another two test messages. Unless you tell me precisely what messages were involved, it is very time-consuming to track this down. However, before you do this read the rest of this message. There are a number of items like, e.g., crossposting to multiple newsgroups, which can sometimes give rise to unexpected behavior between the mailing lists and newsgroups. If the messages in question were *only* sent to the 7tms_r mail address or *only* sent to bionet.molbio.proteins.7tms_r and did not show up in both media, then I would be concerned. There can also be substantial differences in delivery time between news and mail to the same site (more below). > In January, many "Mailing List" people asked the reader of the > "Newsgroup" to stop any discussion about "NutraSweet", but only > "Mailing List" people can read it, "Newsnet" people cannot. I saw discussions about Nutrasweet in both our newsreader and mail archives here. I have not attempted to match them all up one-to-one and instead am simply posting two test messages via news and mail to test the group. > Recently I have more than 10 mails from "Mailing List" people, but > they are totally not appeared in the "NewsNet". What happens? > Dr.Cornelius Krasel and so many people recognized it. It could be that your news feed is convoluted and that it might take several days for the news articles to get to you via other sites while the mail distribution goes from here to you directly. It is very important to look at distribution headers carefully and determine a typical delivery time from our site to yours before assuming that the system is broken. news distribution and mail distribution proceed by entirely different routes. > Do you kindly check it and correct the system? I just posted a test via mail and also via news. If I find out that anything is really broken, I will fix it, but, as noted above, there are other possible sources which might account for your observations. I am ccing the newsgroup on this in case others have the same question. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!bionet.ig.com!not-for-mail From: kristoff@bionet.ig.com (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: test of bionet.molbio.proteins.7tms_r Date: 6 Feb 1995 16:58:09 -0800 Organization: BIOSCI International Newsgroups for Biology Lines: 3 Distribution: world Message-ID: <3h6gj1$n7b@bionet.ig.com> NNTP-Posting-Host: bionet.ig.com Test of the bionet.molbio.proteins.7tms_r USENET newsgroup and the news-to-mail gateway. From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!BIONET.IG.COM!kristoff From: kristoff@BIONET.IG.COM (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: test of 7tms_r@net.bio.net Date: 6 Feb 1995 16:56:32 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net test of 7tms_r@net.bio.net distribution, please ignore. From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: UNSUBSCRIBING, BIOSCI ARCHIVES, ADDRESS DATABASE & BIOSCI FAQ Date: 6 Feb 1995 17:26:23 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 338 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net Four important items follow: How to cancel e-mail subscriptions to BIOSCI newsgroups, BIOSCI archive searching, the BIOSCI FAQ, and the BIOSCI User Address Directory form. If you have not yet listed yourself in our BIOSCI user directory, please take a few minutes to complete and return the form below. If your personal information has changed since you listed yourself, please send us a complete new updated form. We can not make manual revisions to existing entries. 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Note that the "e-mail network: " line below is for specifying, e.g., "Internet," "BITNET," "EARN," "JANET," or whatever other network that your computer may be on. If you are uncertain about any field, please feel free to leave it blank, but please DO NOT DELETE the field identifier from the form! In the first field below, "New information or Update ...", please enter "N" if this is the first time that you have registered in the directory or "U" if you are correcting a listing that you sent to us previously. The comment: lines may be used for anything that you like but PLEASE DO NOT DELETE THEM FROM THE FORM OR ALTER THEM. One suggested use is to list the names of the newsgroups in which you participate. Please use the MAILING LIST name (see below - the latest version of the list can be requested from biosci@net.bio.net) instead of the USENET name even if you don't participate by e-mail. WAIS might get confused by the periods in the USENET names. This allows one to retrieve via WAIS or waismail the list of participants in a particular group. For example: comment: ARABIDOPSIS PLANT-BIOLOGY BIONEWS On the comment: lines use these names below ---- NOT the USENET names below MAILING LIST NAME USENET Newsgroup Name ----------------- --------------------- ACEDB-SOFT bionet.software.acedb AGEING bionet.molbio.ageing AGROFORESTRY bionet.agroforestry ARABIDOPSIS bionet.genome.arabidopsis ASCB bionet.prof-society.ascb BIOCAN bionet.prof-society.cfbs BIOFORUM bionet.general BIO-INFORMATION-THEORY bionet.info-theory BIONAUTS bionet.users.addresses BIONEWS bionet.announce BIO-JOURNALS bionet.journals.contents BIO-MATRIX bionet.molbio.bio-matrix BIOPHYSICAL-SOCIETY bionet.prof-society.biophysics BIOPHYSICS bionet.biophysics BIO-SOFTWARE bionet.software BIOTHERMOKINETICS bionet.metabolic-reg BIO-WWW bionet.software.www CARDIOVASCULAR-RESEARCH bionet.biology.cardiovascular CELEGANS bionet.celegans CELL-BIOLOGY bionet.cellbiol CHLAMYDOMONAS bionet.chlamydomonas CHROMOSOMES bionet.genome.chromosomes COMPUTATIONAL-BIOLOGY bionet.biology.computational CSM bionet.prof-society.csm CYTONET bionet.cellbiol.cytonet DROSOPHILA bionet.drosophila EMBL-DATABANK bionet.molbio.embldatabank EMF-BIO bionet.emf-bio EMPLOYMENT bionet.jobs EMPLOYMENT-WANTED bionet.jobs.wanted FASEB bionet.prof-society.faseb GDB bionet.molbio.gdb GENBANK-BB bionet.molbio.genbank GENETIC-LINKAGE bionet.molbio.gene-linkage GRASSES-SCIENCE bionet.biology.grasses HIV-MOLECULAR-BIOLOGY bionet.molbio.hiv HUMAN-GENOME-PROGRAM bionet.molbio.genome-program IMMUNOLOGY bionet.immunology INFO-GCG bionet.software.gcg JOURNAL-NOTES bionet.journals.note METHODS-AND-REAGENTS bionet.molbio.methds-reagnts MICROBIOLOGY bionet.microbiology MOLECULAR-EVOLUTION bionet.molbio.evolution MOLECULAR-MODELLING bionet.molec-model MOLLUSC-MOLECULAR-NEWS bionet.molbio.molluscs MYCOLOGY bionet.mycology NEUROSCIENCE bionet.neuroscience N2-FIXATION bionet.biology.n2-fixation PARASITOLOGY bionet.parasitology PHOTOSYNTHESIS bionet.photosynthesis PLANT-BIOLOGY bionet.plants POPULATION-BIOLOGY bionet.population-bio PROTEIN-ANALYSIS bionet.molbio.proteins PROTEIN-CRYSTALLOGRAPHY bionet.xtallography PROTISTA bionet.protista RAPD bionet.molbio.rapd SCIENCE-RESOURCES bionet.sci-resources STADEN bionet.software.staden STRUCTURAL-NMR bionet.structural-nmr TROPICAL-BIOLOGY bionet.biology.tropical URODELES bionet.organisms.urodeles VIROLOGY bionet.virology WOMEN-IN-BIOLOGY bionet.women-in-bio YEAST bionet.molbio.yeast ZBRAFISH bionet.organisms.zebrafish Listing newsgroups on the comment: line is optional, of course. Thanks again for your cooperation! --------------- please cut here and return portion below --------------- New information or Update to old record (enter N or U): date (DD-MM-YY): first name: middle initial: family name: job title: e-mail address: e-mail network: phone number: FAX number: institution: address1: address2: address3: city: state/province: country: postal code: research interest: research interest: comment: comment: comment: comment: comment: From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!biosci!not-for-mail From: kristoff@net.bio.net (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 6 Feb 1995 17:12:26 -0800 Organization: BIOSCI International Newsgroups for Biology Lines: 16 Distribution: world Message-ID: <3h6hdq$r12@net.bio.net> References: NNTP-Posting-Host: net.bio.net In article , BIOSCI Administrator wrote: >We tested the system previously and it worked fine. I have just >posted another two test messages. Both of the two tests just posted showed up in both the news and mail system here. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!glaxo.com!brown~pj From: brown~pj@glaxo.com ("Peter J. Brown") Newsgroups: bionet.molbio.proteins.7tms_r Subject: unsubscaribe Date: 6 Feb 1995 10:06:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: <34307160205991.591343@USA> NNTP-Posting-Host: net.bio.net [This message is converted from WPS-PLUS to ASCII] unsubscribe From owner-7tms_r@net.bio.net Sun Feb 05 22:00:00 1995 Path: biosci!WELCHLINK.WELCH.JHU.EDU!mlevine From: mlevine@WELCHLINK.WELCH.JHU.EDU (MICHAEL A LEVINE) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: membrane spanning domains Date: 6 Feb 1995 13:56:13 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: References: NNTP-Posting-Host: net.bio.net the receptor for calcitonin gene related protein (CGRP) activates adenylyl cyclase, most likely via Gs, but the receptor is not 7-tms; also, IGF-2 receptor can activate g proteins. On Mon, 6 Feb 1995, AJ. Collier wrote: > > Does anyone know of a G-protein linked receptor which doesnt possess 7 membrane spanning domains. > Thanks > Adam Collier > :quit > > > > From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!cs.utexas.edu!uunet!CS.Arizona.EDU!biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 6 Feb 1995 17:08:37 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 57 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net > > Dear Dr.Dave Kristofferson, > > I am interested in G-protein Coupled Receptor, so I join 7tms_r Mailing > list, and also read bionet.molbio.proteins.7tms_r. But it is strange. > Suppose that I post some comments to 7tms_r@net.bio.net, and it is > distributed to the "Mailing List" people, but not appeared in the > "Bionet.molbio.proteins.7tms_r" Newsgroup. We tested the system previously and it worked fine. I have just posted another two test messages. Unless you tell me precisely what messages were involved, it is very time-consuming to track this down. However, before you do this read the rest of this message. There are a number of items like, e.g., crossposting to multiple newsgroups, which can sometimes give rise to unexpected behavior between the mailing lists and newsgroups. If the messages in question were *only* sent to the 7tms_r mail address or *only* sent to bionet.molbio.proteins.7tms_r and did not show up in both media, then I would be concerned. There can also be substantial differences in delivery time between news and mail to the same site (more below). > In January, many "Mailing List" people asked the reader of the > "Newsgroup" to stop any discussion about "NutraSweet", but only > "Mailing List" people can read it, "Newsnet" people cannot. I saw discussions about Nutrasweet in both our newsreader and mail archives here. I have not attempted to match them all up one-to-one and instead am simply posting two test messages via news and mail to test the group. > Recently I have more than 10 mails from "Mailing List" people, but > they are totally not appeared in the "NewsNet". What happens? > Dr.Cornelius Krasel and so many people recognized it. It could be that your news feed is convoluted and that it might take several days for the news articles to get to you via other sites while the mail distribution goes from here to you directly. It is very important to look at distribution headers carefully and determine a typical delivery time from our site to yours before assuming that the system is broken. news distribution and mail distribution proceed by entirely different routes. > Do you kindly check it and correct the system? I just posted a test via mail and also via news. If I find out that anything is really broken, I will fix it, but, as noted above, there are other possible sources which might account for your observations. I am ccing the newsgroup on this in case others have the same question. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!cs.utexas.edu!uunet!CS.Arizona.EDU!biosci!bionet.ig.com!not-for-mail From: kristoff@bionet.ig.com (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: test of bionet.molbio.proteins.7tms_r Date: 6 Feb 1995 16:58:09 -0800 Organization: BIOSCI International Newsgroups for Biology Lines: 3 Distribution: world Message-ID: <3h6gj1$n7b@bionet.ig.com> NNTP-Posting-Host: bionet.ig.com Test of the bionet.molbio.proteins.7tms_r USENET newsgroup and the news-to-mail gateway. From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!howland.reston.ans.net!swrinde!cs.utexas.edu!uunet!CS.Arizona.EDU!biosci!BIONET.IG.COM!kristoff From: kristoff@BIONET.IG.COM (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: test of 7tms_r@net.bio.net Date: 6 Feb 1995 16:56:32 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net test of 7tms_r@net.bio.net distribution, please ignore. From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!cs.utexas.edu!uunet!CS.Arizona.EDU!biosci!biosci!not-for-mail From: kristoff@net.bio.net (David Kristofferson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 6 Feb 1995 17:12:26 -0800 Organization: BIOSCI International Newsgroups for Biology Lines: 16 Distribution: world Message-ID: <3h6hdq$r12@net.bio.net> References: NNTP-Posting-Host: net.bio.net In article , BIOSCI Administrator wrote: >We tested the system previously and it worked fine. I have just >posted another two test messages. Both of the two tests just posted showed up in both the news and mail system here. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!galaxy.ucr.edu!ratatosk.yggdrasil.com!news.duke.edu!solaris.cc.vt.edu!news.bluesky.net!udel!news.sprintlink.net!pipex!lyra.csx.cam.ac.uk!warwick!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (AJ. Collier) Subject: membrane spanning domains Message-ID: Sender: usenet@info.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: zeus.bris.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL2] Date: Mon, 6 Feb 1995 15:39:02 GMT Lines: 7 Does anyone know of a G-protein linked receptor which doesnt possess 7 membrane spanning domains. Thanks Adam Collier :quit From owner-7tms_r@net.bio.net Mon Feb 06 22:00:00 1995 Path: biosci!AMYLIN.COM!keith_albrandt From: keith_albrandt@AMYLIN.COM ("Keith Albrandt") Newsgroups: bionet.molbio.proteins.7tms_r Subject: FWD>RE>membrane spanning do Date: 7 Feb 1995 10:14:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 49 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Mail*Link(r) SMTP FWD>RE>membrane spanning dom What's the evidence that the receptor for CGRP is not a 7tm? Has it been cloned? Thanks. Keith Albrandt ------------------------------ Date: 2/5/95 10:21 PM From: MICHAEL A LEVINE the receptor for calcitonin gene related protein (CGRP) activates adenylyl cyclase, most likely via Gs, but the receptor is not 7-tms; also, IGF-2 receptor can activate g proteins. On Mon, 6 Feb 1995, AJ. Collier wrote: > > Does anyone know of a G-protein linked receptor which doesnt possess 7 membrane spanning domains. > Thanks > Adam Collier > :quit > > > > ------------------ RFC822 Header Follows ------------------ Received: by amylin.com with SMTP;6 Feb 1995 15:36:10 -0800 Received: from net.bio.net by jason (4.1/SMI-4.0) id AA25456; Mon, 6 Feb 95 15:30:24 PST Received: (from daemon@localhost) by net.bio.net (8.6.9/8.6.6) id NAA16841 for 7tms_r-list; Mon, 6 Feb 1995 13:56:25 -0800 Received: (from news@localhost) by net.bio.net (8.6.9/8.6.6) id NAA16831 for 7tms_r-arpanet; Mon, 6 Feb 1995 13:56:22 -0800 To: 7tms_r@net.bio.net From: mlevine@welchlink.welch.jhu.edu (MICHAEL A LEVINE) Subject: Re: membrane spanning domains Date: 6 Feb 1995 13:56:13 -0800 Sender: daemon@net.bio.net Message-Id: Nntp-Posting-Host: net.bio.net From owner-7tms_r@net.bio.net Tue Feb 07 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!howland.reston.ans.net!ix.netcom.com!netcom.com!graul From: graul@netcom.com (Rick Graul) Subject: Re: membrane spanning domains Message-ID: Sender: graul@netcom.com (Rick Graul) Reply-To: graul@netcom.com (Rick Graul) Organization: NETCOM On-line Communication Services (408 261-4700 guest) References: Date: Thu, 9 Feb 1995 03:58:06 GMT Lines: 12 dhajc@zeus.bris.ac.uk (AJ. Collier) writes: >Does anyone know of a G-protein linked receptor which doesnt possess 7 membrane spanning domains. The GPCR homolog vc03_sfvka, from the shope fibroma virus, appears to be missing all of transmembrane domain 1 and the beginning of 2. Rick -- ********************************************************************** * Rick Graul internet: graul@netcom.com * ********************************************************************** From owner-7tms_r@net.bio.net Tue Feb 07 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!cs.utexas.edu!convex!news.duke.edu!news-server.ncren.net!concert!hearst.acc.Virginia.EDU!murdoch!usenet From: Bruce Gaylinn Subject: Re: FWD>RE>membrane spanning do X-Nntp-Posting-Host: bootp-205-42.bootp.virginia.edu Message-ID: Sender: usenet@murdoch.acc.Virginia.EDU Organization: University of Virginia References: Date: Wed, 8 Feb 1995 18:38:12 GMT Lines: 32 keith_albrandt@AMYLIN.COM ("Keith Albrandt") wrote: > > Mail*Link(r) SMTP FWD>RE>membrane spanning dom > > What's the evidence that the receptor for CGRP is not a 7tm? Has it been > cloned? > Thanks. > > Keith Albrandt > > > ------------------------------ > Date: 2/5/95 10:21 PM > From: MICHAEL A LEVINE > > the receptor for calcitonin gene related protein (CGRP) activates > adenylyl cyclase, most likely via Gs, but the receptor is not 7-tms; > also, IGF-2 receptor can activate g proteins. > I too am surprised at the suggestion that the CGRP receptor is not a 7-tmr. 1).CGRP has strong sequence similarity to calcitonin which does have a 7tmr. 2). CGRP is coupled to cAMP and GTP shifts the high affinity site to a lower affinity state. 3). CGRP cross- linking suggests a receptor with size and glycosylation typical of a 7tmr. Am I miss-reading the literature, is there a new surprising clone, or is this some sort of missunderstanding? From; Bruce Gaylinn bg2g@virginia.edu From owner-7tms_r@net.bio.net Tue Feb 07 22:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 8 Feb 1995 10:56:10 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 113 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net > Dr. Kristofferson, > > I think that there is some propagation problem between net.bio.net and > for example this site, thereby I cannot read 7tms via news. I am > not familiar enough with news to explain or resolve this problem. > I asked the local news admin about it and he said during the creation > of the newsgroup there was a problem with the email address of the > "responsible person". Maybe this has caused troubles further upstream > (i.e. between net.bio.net and news.biochem.mpg.de). > > Maybe it would be helpful if people mailed replies to your test message > (only if read by news, of course) to track the culprit down. If it is of > help, I can email you a header of an established group, such as > bionet.announce. > > This problem occurs here not only with bionet.molbio.proteins.7tms, but > with any lately created bionet group. > > Hope that clears things up a bit, > --Cornelius Krasel This problem is something that we have had to contend with increasingly as USENET gets flooded with trash in the alt domain and sites turn off automatic newsgroup creation. This means that at each site the newsgroup administrator has to act *manually* on newsgroup creation messages. Valuable ones can get lost in the general flow of garbage on the net. You should talk to your administrator about ways of ensuring that all bionet newgroup messages get acted upon at your site, but your administrator can also use our "checkgroups" message to get caught up. I enclose the relevant portion of our BIOSCI FAQ below. Please be aware that your news administrator is probably harried by other tasks and will not do these things unless you call them to his/her attention. I am ccing the 7tms_r group on this too. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net ---------------------------------------------------------------------- I HEAR THAT A NEW BIONET NEWSGROUP WAS CREATED BUT WHY ISN'T IT AT MY SITE YET? This could happen for a variety of reasons. When we create a new Usenet newsgroup, a "newgroup" message is sent out from net.bio.net to news administrators at Usenet sites around the world. Many sites are configured so that such newgroup messages are acted upon automatically and the group is established without human intervention. However, due to the growing volume of Usenet newsgroups, many sites have turned off automatic newgroup creation and require human intervention to create a new Usenet group in response to a newgroup message. If you know from reading BIONEWS/bionet.announce or from contacting the BIOSCI staff at biosci-help@net.bio.net that a new bionet newsgroup should be in existance, please contact your local news administrator and ask them if they acted on the newgroup message. Newgroup messages can sometimes be simply overlooked by the news administrator and sometimes they may not be received if a Usenet site upstream on the net from you had a problem and did not pass on the message. Please let your news adminstrator know that a bionet "checkgroup" message is posted on the first of each month to bionet.announce. Your news administrator can use the contents of that message to update your local list of bionet Usenet newsgroups. Please note that all new BIOSCI/bionet newsgroups are announced on BIONEWS/bionet.announce as soon as they are ready for use. If you see this announcement, this means that the BIOSCI staff has tested the group and is (1) sure that it works, and (2) knows that it has propagated to at least some other sites in both North America and Europe. We obviously can not check for propagation to the thousands of sites on Usenet. Depending upon your source of Usenet news, there may be a delay of several days before the newgroup message reaches your site. The announcement of the newsgroup availability, however, is always sent to bionet.announce *after* the newgroup message has been sent and after system tests have been run. ---------------------------------------------------------------------- A NEW BIONET USENET GROUP HAS BEEN CREATED AT MY SITE BUT THERE ARE NO MESSAGES IN IT. HOWEVER, I SEE THAT MESSAGES ARE BEING SENT OUT TO THE MAILING LIST. WHY DO THE CONTENTS DIFFER? If you experience the problem above, please contact your local news administrator and have them check with the site that sends you your bionet Usenet news feed. We explained in a question above that new Usenet newsgroups are created in response to a "newgroup" message which is sent out to all Usenet news administrators. It is possible that your news administrator acted on the message to create the group, but that the site which sends you bionet Usenet news did not. Having your Usenet news administrator contact the administrator at the upstream site can usually resolve the problem. If you have a problem getting a reliable bionet Usenet news feed, please contact biosci-help@net.bio.net. ---------------------------------------------------------------------- HOW DO I REQUEST OR CANCEL E-MAIL SUBSCRIPTIONS TO BIOSCI NEWSGROUPS? Instructions for subscribing and unsubscribing to the BIOSCI newsgroups by e-mail are included in the BIOSCI info sheet which can be obtained from either of the following two addresses: Administrative Address Location ---------------------- -------- biosci@daresbury.ac.uk Europe, Africa, and Central Asia biosci@net.bio.net Americas and the Pacific Rim ---------------------------------------------------------------------- From owner-7tms_r@net.bio.net Wed Feb 08 22:00:00 1995 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news.sprintlink.net!uunet!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: biobone@aol.com (Bio Bone) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor density and transfection Date: 9 Feb 1995 18:44:57 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 9 Sender: root@newsbf02.news.aol.com Message-ID: <3he9dp$7rg@newsbf02.news.aol.com> References: <199502091026.CAA21272@net.bio.net> Reply-To: biobone@aol.com (Bio Bone) NNTP-Posting-Host: newsbf02.mail.aol.com We have a number of CHO clones that have been transfected with the thrombin receptor. We have determined that these clones generally are expressing between 100,000 and 150,000 receptors/cell. Over time, we have noticed that the expression level has generally gone down only slightly although there is one clone that is will lose expression after many passages. Bradley Bone COR Therapeutics, Inc. From owner-7tms_r@net.bio.net Wed Feb 08 22:00:00 1995 Path: biosci!NET.BIO.NET!biosci-help From: biosci-help@NET.BIO.NET (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMS_R "Newsnet" and "Mailing List" Date: 9 Feb 1995 10:22:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 55 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: biosci-help@net.bio.net NNTP-Posting-Host: net.bio.net > > It might be something else as well. I get the newsgroup but also continue > to read as email. It seems to me that only some of the stuff that I see > in email makes it to the newsgroup. I can't be sure of this since I > haven't been paying particularly close attention but that is my impression. > > Max > I am absolutely confident that messages posted to the two BIOSCI sites get distributed both by mail and news. A number of issues can develop downstream at other news sites and can cause the kind of discrepancy mentioned above. I'll comment on the most likely cause: If your site rapidly expires the 7tms_r newsgroup (perhaps they set your news software to delete all messages older than one day - this is a locally settable parameter), you may miss messages in the news system if you do not read it religiously. News users should ask there local systems administrators to set the expiration date on the bionet newsgroups to a period which corresponds with the frequency of their access. The bionet groups are not high volume so this should not create disk space problems on your machine. I'm sure after reading these various posts many of you are starting to think that you should only participate by e-mail. Personally, I think that e-mail is the worst possible option and a horrendous waste of your time **IF** you have a properly configured and working newsreader. If you do not, then you are stuck with having to get all of the messages to the newsgroup dumped in your mail file. I don't know about the rest of you, but if I had to read a part of each article in a journal before I could decide whether or not it was of interest, I would quickly conclude that the journals were full of junk that wasted my time. Everyone reads journals by first skimmimg the table of contents and selecting the small number of articles of interest in each issue. A newsreader sorts the messages by discussion topic, thus automatically creating a table of contents for you, and lets you quickly skim what is being discussed. You don't have to delete anything out of your mail file that is not of interest. You can quickly skip SUBSCRIBE messages and other off-the-topic or inappropriate postings without having to delete them. It's usually e-mail users that get bent out of shape about chain letters, etc. on the net. News users rarely even look at them. Hopefully the above will convince you that, although it may take some effort to talk to your news administrator now and get them to configure the newsgroups properly for you, it will over the long run save you a tremendous ammount of time and effort. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Wed Feb 08 22:00:00 1995 Path: biosci!VM.CNUCE.CNR.IT!TMWERGE%IRMISS.BITNET From: TMWERGE%IRMISS.BITNET@VM.CNUCE.CNR.IT (Thomas) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor density and transfection Date: 9 Feb 1995 02:28:47 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502091026.CAA21272@net.bio.net> NNTP-Posting-Host: net.bio.net Good morning out-there| I have only recently joint this list and realize that I may be asking a FAQ. Nevertheless, how do one equalize the receptor density per cell in a transient system like COS cells of two differently expressed receptors (generally a high wildtype and a low mutant)? I immagine that one may vary the DNA conc of the be st expressed receptor (adding up empthy vector DNA so that the conc of DNA rema ins the unaltered). Would such an approach, given it yeilds equal receptor conc per well, insure that also the receptor density per cell is comparable. And talking about receptor density, has anybody examined the distribution of re ceptor densities in transfected COS cells; i.e. is it accually true that COS cells, independently of number of introduced plasmids, amplify the plasmid DNA as to reach a common identical (or very similar) copy number per cell, whic h would imply an "all or nothing" type of expression? A second issue: how stable are stable CHO clones. Has anybody experience which how (or if) receptor expression varies in such clones, either with passage of t he clone (which I guess is expected) or with its actual state of growth? Have a nice day| Cheers Thomas ››››››››››››››››››››››››››››››››› THOMAS M. WERGE I.S.S. LABORATORY OF PHARMACOLOGY VIALE REGINA ELENA 299 - 00161 ROME ITALY TEL. (+39) 6 4990 2386 - FAX (+39) 6 444 0053 E-MAIL TMWERGE @ ISS.IT From owner-7tms_r@net.bio.net Thu Feb 09 22:00:00 1995 Path: biosci!MSMAIL.BMS.COM!Carlson_Kenneth_E.PRILVMS3 From: Carlson_Kenneth_E.PRILVMS3@MSMAIL.BMS.COM (Carlson Kenneth E) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 10 Feb 1995 10:43:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: subscribe From owner-7tms_r@net.bio.net Sat Feb 11 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!cs.utexas.edu!swrinde!gatech!newsfeed.pitt.edu!uunet!zib-berlin.de!informatik.tu-muenchen.de!lrz-muenchen.de!ipp-garching.mpg.de!alf.biochem.mpg.de!krasel From: krasel@alf.biochem.mpg.de (Cornelius Krasel) Newsgroups: bionet.xtallography,bionet.molbio.proteins.7tms_r Subject: Re: Integral Membrane Proteins? Date: 11 Feb 1995 12:42:37 GMT Organization: Rechenzentrum der Max-Planck-Gesellschaft in Garching Lines: 50 Distribution: world Message-ID: <3hibbt$1e00@sat.ipp-garching.mpg.de> References: <3he93u$5hp@emory.mathcs.emory.edu> NNTP-Posting-Host: alf2.biochem.mpg.de X-Newsreader: TIN [version 1.2 PL2] Xref: biosci bionet.xtallography:1521 bionet.molbio.proteins.7tms_r:173 [This has been crossposted to bionet.molbio.proteins.7tms_r] T. J. Murphy (medtjm@bimcore.emory.edu) wrote: > My interests include "7TM" receptors coupled to GTP-binding proteins. > Since "7TM" receptors are integral membrane proteins, they have proven > refractory to known approaches for crystallization and structural solution > (as I understand it). 1) It is possible to crystallize integral membrane proteins (see for example the photosynthetic reaction center or the porins). It is, however, much more difficult than crystallizing soluble proteins. 2) It is very difficult (yet) to express G-protein coupled receptors in quantities needed for expression. E. coli are rather reluctant to produce these proteins (you get about five to ten molecules per cell). Baculovirus expression is better, but still not comparable to soluble proteins in baculovirus. In mammalian cells, expression can be forced to rather high levels but here arise the problems of culturing large amounts of cells. 3) There is a projection structure of bovine rhodopsin available by G. Schertler. Bovine rhodopsin can be purified rather easily in large amounts from eyes (thereby circumventing the problems pointed out in #2 above). > if one produces segments of a 7TM receptor protein, each containing a single > predicted membrane spanning region, what is the likelihood that these > segments could be crystallized, and solved individually? I am not sure if this approach has been tried with membrane segments of G-protein coupled receptors, but the PDB contains some NMR structures of membrane-spanning peptides derived from bacteriorhodopsin whose structure has been solved by NMR. It is very likely that indeed the transmembrane structures of G-protein coupled receptors are more or less amphipathic helices, with the more hydrophilic part pointing towards the center of the receptor. Based on this assumption and on mutational data, a model has been constructed by Joyce Baldwin (Baldwin, Curr. Op. Struct. Biol. 6, 180-190, 1994 and references therein) which attempts to fit all the experimental data. Now let the modellers stand up :-) --Cornelius. -- /* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */ /* email: krasel@alf.biochem.mpg.de fax: +49 89 8578 3795 */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Sat Feb 11 22:00:00 1995 Path: biosci!MSMAIL.BMS.COM!Seiler_Steven_M.PRILVMS3 From: Seiler_Steven_M.PRILVMS3@MSMAIL.BMS.COM (Seiler Steven M) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Partial agonists Date: 12 Feb 1995 14:03:43 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net subscribe (again). For those who are interested my new address is: Seiler_Steven_M.PRILVMS3@msmail.bms.com While I've got your attention, I was wondering what's the best current description of the molecular basis for partial agonists? (Literature as well as personal opinions welcome.) Also, what does every one think of using GTPase activity to measure the degree of agonist activity? /Steve Seiler From owner-7tms_r@net.bio.net Sat Feb 11 22:00:00 1995 Path: biosci!MIZAR.USC.EDU!cminkin From: cminkin@MIZAR.USC.EDU ("Cedric Minkin") Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 12 Feb 1995 16:33:00 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Sun Feb 12 22:00:00 1995 Path: biosci!CHET.MEDC.UMN.EDU!germana From: germana@CHET.MEDC.UMN.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 13 Feb 1995 14:06:45 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502132200.AA07245@quinn.medc.umn.edu> NNTP-Posting-Host: net.bio.net I am looking for the program KITSCH by Felsenstein to calculate dendograms. May thanks to pointers. Germana Paterlini Dept. of Medicinal Chemistry Universit of Minnesota Germana@quinn.medc.umn.edu .\] From owner-7tms_r@net.bio.net Sun Feb 12 22:00:00 1995 Path: biosci!VM.CNUCE.CNR.IT!TMWERGE%IRMISS.BITNET From: TMWERGE%IRMISS.BITNET@VM.CNUCE.CNR.IT ("Thomas M. Werge") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Rec. density in COS Date: 13 Feb 1995 07:14:19 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 47 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502131512.HAA06459@net.bio.net> NNTP-Posting-Host: net.bio.net Good afternoon| Last week I asked a few questions regarding transfections in COS and CHO cells and the resulting receptor expression and density. Thanks for the answers| (som e of which I received both directly and through the list: Generally the mailing list took 20 to 40 hr more than direct mailing; is that normal?). I would like to follow up on the COS transfection. From what I understood and n ow understand even better from the answers, the expression of receptors in COS has not been address experimentally; i.e. it is not known in which way expresse d receptors are distributed on COS cells, normal distributed or in an 'all or non' fasion. Less seems to be known about which parameters differ, 'receptor n umber per cell' or 'number of expressing cells' (or both), when two receptors a re compared, such as a good expressing wildtype and a bad expressing mutant. When DNA conc. in transfections are adjusted as to give comparable receptor exp ression per well (or just comparable binding per well) it is not known whether this 'equal' expression is obtained by matching both receptor density and numbe r of positive cells (as one would like to), or by changing one of the parameter s only. Given that G-protein coupling in COS is also a bit difficult to control (e.g. p eptide receptors show high affinity binding whereas cationic amines exhibit low affinity) I cannot help asking how one can trust comparative studies of diffen tially expressed receptors (wt vs mut's), in particular when these experiments address the issue of signal transduction? Just one example: I have heard the following type of reasoning: The IC50's (or Kd's) were ..... and the EC50's ...... The Bmax was between 25% and 85% of the wildtype Bmax. Correcting for this diffent expression we see that this or that mutation affects .... However, in my mind, if the expression profiles differ then also the coupling d iffers, and if that is true is becomes hard to compare such receptors. Then onl y stable clones allow this comparison to be done. Needless to say that I do not want to discredit anybodies work, least of all my own) but the more I try to get into this branch of pharmacology the more diffi culties appears. All the best Thomas ››››››››››››››››››››››››››››››››› THOMAS M. WERGE I.S.S. LABORATORY OF PHARMACOLOGY VIALE REGINA ELENA 299 - 00161 ROME ITALY TEL. (+39) 6 4990 2386 - FAX (+39) 6 444 0053 E-MAIL TMWERGE @ ISS.IT From owner-7tms_r@net.bio.net Sun Feb 12 22:00:00 1995 Path: biosci!IREARN.BITNET!emadtajk From: emadtajk@IREARN.BITNET ("E.TAJKHORSHID") Newsgroups: bionet.molbio.proteins.7tms_r Subject: CONGRESS ANNOUNCEMENT Date: 13 Feb 1995 05:01:38 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 132 Sender: daemon@net.bio.net Distribution: world Message-ID: <0098BEE6.CA299CE0.171@IREARN.BITNET> NNTP-Posting-Host: net.bio.net >> 12th IRANIAN CONGRESS OF PHYSIOLOGY AND PHARMACOLOGY << Organized by the Iran University of Medical Sciences in Collaboration with Iranian Society of Physiology and Pharmacology Tehran (Iran), November 6-9, 1995 ****************************************************************************** Dear Colleague: The organizing committee of the 12th ICPP is honored to invite you to participate actively in this meeting. ****************************************************************************** TOPICS (Code): 1) Adverse Drug Reactions (100) 2) Aging (Gerontology) (101) 3) Autonomic Nervous System (102) 4) Biochemistry (103) 5) Biophysics and Biomedical Engineering (104) 6) Biopharmaceutics and Pharmacokinetics (105) 7) Blood and Blood Components (106) 8) Cancer and Neoplasia (107) 9) Cellular and General Physiology (108) 10) Central Nervous Systenm (109) 11) Chemotherapeutic Agents (110) 12) Clinical Pharmacology and Therapeutics (111) 13) Comparative Physiology (112) 14) Developmental Pharmacology and Toxicology (113) 15) Drug Interaction (114) 16) Endocrine System and Reproduction (115) 17) Environmental and Exercise Physiology (116) 18) Enzymes and Enzyme Regulation (117) 19) Ethnopharmacology (118) 20) Gastrointestinal Pharmacology (119) 21) Genetics (120) 22) Heart and Circulation (121) 23) Immunology and Immunopharmacology (122) 24) Inflamnmation and Anti-Inflammatory Agents (123) 25) Local Hormones and Autocoides (124) 26) Membrane and Transport (125) 27) Nutrition (126) 28) Pathobiology (127) 29) Pharmaceutical Chemistry (128) 30) Pharmacognosy (129) 31) Receptor and Molecular Pharmacology (130) 32) Renal Physiology and Pharmacology (131) 33) Respiratory System (132) 34) Toxicology (133) 35) Veterinary Physiology and Pharmacology (134) ****************************************************************************** Free communications will be presented solely as poster presentation. Abstract deadline: MAY 31, 1995 Deadline for abstract submission: April 30, 1995 Language: English / Farsi ------------------------------------------------------------------------------ Registration Form Title: Prof/Dr/Mr/Mrs/Miss/Ms Complete Name: Sex: Degree: Institutional Affiliation: Position Held: Mailing Address: Presenting: Poster ( ) Seminar ( ) Workshop ( ) None ( ) Category Code as appeared in the topic list: 1st. Choice [ ] 2nd Choice [ ] Registration Fees: Prior to June 31, 95 300 US$ ( ) After June 31, 95 400 US$ ( ) Student* 150 US$ ( ) * Please enclose proof of your student status. Payments: Please draft the registration fee, payable to: Prof. Massoud Mahmoudian, Secretary of 12th ICPP, in Bank Melli Iran, Eskan branch, Code 271, Tehran,IRAN and mail the receipt with the registration form to the congress secretary. ------------------------------------------------------------------------------ Just for visa applicants: Father's name: Date and place of birth: Nationality: Occupation: Passport No.: Date & place of issue: Exp. date: Place where visa to be issued: Duration of staying in IRAN: ////////////////////////////////////////////////////////////////////////////// ------------------------------------------------------------------------------ You may fill and email the registration form (and your abstract) to: MASMAH73@IREARN.BITNET ------------------------------------------------------------------------------ For further information contact: Prof. M. Mahmoudian Secretary of the Congress, International Relations Dept., Iran Universioty of Medical Sciences, P.O.Box 15875/6171, Tehran, IRAN FAX: +98-21-8016207 E-MAIL: MASMAH73@IREARN.BITNET From owner-7tms_r@net.bio.net Sun Feb 12 22:00:00 1995 Path: biosci!gwdg.de!fwuerri From: fwuerri@gwdg.de (Frank Wuerriehausen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: unsubscribe Date: 13 Feb 1995 22:46:54 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502140646.AA28049@gwdu03.gwdg.de> NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Mon Feb 13 22:00:00 1995 Path: biosci!bms.com!PARKER_E From: PARKER_E@bms.com Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoctoral Position Date: 14 Feb 1995 09:28:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HN1FCXIQFW90N3G4@bms.com> NNTP-Posting-Host: net.bio.net I am looking for a postdoctoral fellow to work in my laboratory here at Bristol-Myers Squibb Company. The postdoctoral research will focus on one or more of the following areas: 1. Identification, characterization and cloning of peptides and peptide receptors involved in the central regulation of feeding behavior and energy metabolism. 2. Molecular characterization of nutrient sensing mechanisms in the brain. 3. Identification of proteins that are differentially expressed in the brains of lean vs. obese animals. Ideally, I would like to find someone with experience in all common molecular biology techniques. Experience in cDNA library construction and screening, expression cloning, site-directed mutagenesis, in situ hybridization, immunocytochemistry, radioligand binding and/or signal transduction assays would be particularly helpful. If you know of any interested persons, please have them forward their CVs to Eric M. Parker Dept. of CNS Biology, Dept. 404 Bristol-Myers Squibb Co. 5 Research Parkway Wallingford, CT 06492 Phone: (203)284-6720 Fax: (203)284-7569 Thanks alot for your help! Eric From owner-7tms_r@net.bio.net Mon Feb 13 22:00:00 1995 Path: biosci!rutgers!csn!ub!netfs.dnd.ca!dgbt!nott!torn!howland.reston.ans.net!cs.utexas.edu!uunet!in1.uu.net!caen!zip.eecs.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Rec. density in COS Date: 14 Feb 1995 16:18:07 GMT Organization: University of Michigan Lines: 24 Message-ID: <3hql3v$14k@lastactionhero.rs.itd.umich.edu> References: <199502131512.HAA06459@net.bio.net> NNTP-Posting-Host: warbler.med.umich.edu TMWERGE%IRMISS.BITNET@VM.CNUCE.CNR.IT ("Thomas M. Werge") wrote: > However, in my mind, if the expression profiles differ then also the coupling d > iffers, and if that is true is becomes hard to compare such receptors. Then onl > y stable clones allow this comparison to be done. > > Needless to say that I do not want to discredit anybodies work, least of all my > own) but the more I try to get into this branch of pharmacology the more diffi > culties appears. Thomas, You have pointed out many of the difficulties with transient transfections. They are particularly difficult for any quantitative comparison of responses which is why many people take the extra effort to create stable lines. It is probably more appropriate to draw qualitative conclusions from transients - this works and this doesn't. Also if there is a huge difference between two receptors then it is likely to be telling you something. The better you do with transfection efficiency the less problem you are likely to have from transients. If the transfection efficiency varies from 5-25% it is much more of a problem than if it varies from 70-90%. Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Mon Feb 13 22:00:00 1995 Path: biosci!nlm.nih.gov!cosmides From: cosmides@nlm.nih.gov Newsgroups: bionet.molbio.proteins.7tms_r Subject: unsubscribe Date: 14 Feb 1995 04:36:03 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502140735.21813.AA@occshost.nlm.nih.gov> NNTP-Posting-Host: net.bio.net UNSUBSCRIBE From owner-7tms_r@net.bio.net Mon Feb 13 22:00:00 1995 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!howland.reston.ans.net!news.sprintlink.net!news.bluesky.net!usenet.eel.ufl.edu!gmi!msunews!caen!zip.eecs.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Rec. density in COS Date: 14 Feb 1995 16:17:48 GMT Organization: University of Michigan Lines: 24 Message-ID: <3hql3d$pj@lastactionhero.rs.itd.umich.edu> References: <199502131512.HAA06459@net.bio.net> NNTP-Posting-Host: warbler.med.umich.edu TMWERGE%IRMISS.BITNET@VM.CNUCE.CNR.IT ("Thomas M. Werge") wrote: > However, in my mind, if the expression profiles differ then also the coupling d > iffers, and if that is true is becomes hard to compare such receptors. Then onl > y stable clones allow this comparison to be done. > > Needless to say that I do not want to discredit anybodies work, least of all my > own) but the more I try to get into this branch of pharmacology the more diffi > culties appears. Thomas, You have pointed out many of the difficulties with transient transfections. They are particularly difficult for any quantitative comparison of responses which is why many people take the extra effort to create stable lines. It is probably more appropriate to draw qualitative conclusions from transients - this works and this doesn't. Also if there is a huge difference between two receptors then it is likely to be telling you something. The better you do with transfection efficiency the less problem you are likely to have from transients. If the transfection efficiency varies from 5-25% it is much more of a problem than if it varies from 70-90%. Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Wed Feb 15 22:00:00 1995 Path: biosci!VM.CNUCE.CNR.IT!TMWERGE%IRMISS.BITNET From: TMWERGE%IRMISS.BITNET@VM.CNUCE.CNR.IT ("Thomas M. Werge") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Is the mail server down? Date: 16 Feb 1995 07:26:45 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502161526.HAA16121@net.bio.net> NNTP-Posting-Host: net.bio.net I send the list a mail on Mon. Feb. 13 which has not yet made it back here. Sin ce I did get a reply to the mail from Neubig (thanks a lot|) my message most be out there somewhere. Other 7tm'ers in Italy didn't received my mail either. Is some link down or is the served overworked? All the best Thomas ››››››››››››››››››››››››››››››››› THOMAS M. WERGE I.S.S. LABORATORY OF PHARMACOLOGY VIALE REGINA ELENA 299 - 00161 ROME ITALY TEL. (+39) 6 4990 2386 - FAX (+39) 6 444 0053 E-MAIL TMWERGE @ ISS.IT From owner-7tms_r@net.bio.net Thu Feb 16 22:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!gatech!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins,bionet.molbio.proteins.7tms_r Subject: Re: G-proteins Date: 17 Feb 1995 11:18:13 GMT Organization: University of Michigan Lines: 46 Message-ID: <3i20ll$b76@lastactionhero.rs.itd.umich.edu> References: <3hud18$qff@bigboote.WPI.EDU> NNTP-Posting-Host: pm002-16.dialip.mich.net Xref: biosci bionet.molbio.proteins:3745 bionet.molbio.proteins.7tms_r:185 hammer@wpi.WPI.EDU (Paul Eugene Hammerstrom) wrote: > > I'm working on a paper and I need to find 10 active/interactive sites > of G-protein subunits, the properties of each site, and the result of > that interaction. Anyone's help would be much appreciated. Paul, There have been 4 papers in the last year on xtal structure of G protein alpha subunits that will have much useful information: 1. Noel, J. P., H. E. Hamm, and P. B. Sigler. 1993. The 2.2 Å crystal structure of transducin-a complexed with GTP-gamma-S. Nature 366:654-663. 2. Lambright, D. G., J. P. Noel, H. E. Hamm, and P. B. Sigler. 1994. Structural determinants for activation of the a-subunit of a heterotrimeric G protein. Nature 369:621-628. 3. Another in Nature from this same group more recently on GDP-AlF bound transducin. 4. Sprang/Gilman lab collaboratin also in Nature on several mutant G alpha-i's. You might also want to look at a review by Conklin and Bourne in Cell last year: 1. Conklin, B. R. and H. R. Bourne. 1993. Structural elements of Ga subunits that interact with Gbt, receptors, and effectors. Cell 73:631-641. We also had a paper in JBC recently on a receptor-peptide interaction site. One of the interesting results was that there was a significant interaction with beta subunit which we are trying hard to map. 1. Taylor, J. M., G. Jacob-Mosier, R. G. Lawton, A. E. Remmers, and R. R. Neubig. 1994. Binding of an a2-adrenergic receptor peptide to Gb and the N-terminus of Ga. J Biol. Chem. 269:27618-27624. There is also a newsgroup specifically devoted to G protein coupled receptors that you might want to look into. bionet.molbio.proteins.7tms_r Good luck. Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Thu Feb 16 22:00:00 1995 Path: biosci!HLSUN.RED-CROSS.ORG!hlatim From: hlatim@HLSUN.RED-CROSS.ORG ("timothy hla") Newsgroups: bionet.molbio.proteins.7tms_r Subject: G-protein / receptor interaction Date: 17 Feb 1995 06:44:48 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502171440.AA18740@hlsun.red-cross.org> Reply-To: "timothy hla" NNTP-Posting-Host: net.bio.net Dear g-ologists: I have perhaps a niave question. What does the affinity of the 3rd loop of the G-protein-coupled receptors (determined by physical association of recombinant third loop fusion protein to native Galphas ) to a specific G protein alpha subunit mean in terms of productive signalling ? Thanks Tim Hla Timothy Hla, Ph.D. Scientist II Department of Molecular Biology Holland Laboratory, American Red Cross 15601 Crabbs Branch Way Rockville, MD 20855, USA ph: 301-738-0567 fx: 301-738-0465 e-mail: hlatim@hlsun.red-cross.org From owner-7tms_r@net.bio.net Thu Feb 16 22:00:00 1995 Path: biosci!EMBL-HEIDELBERG.DE!Gert.Vriend From: Gert.Vriend@EMBL-HEIDELBERG.DE Newsgroups: bionet.molbio.proteins.7tms_r Subject: TM7 server update Date: 17 Feb 1995 08:06:14 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 90 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HN5W7P8GHEIIQZXA@EMBL-Heidelberg.DE> NNTP-Posting-Host: net.bio.net Dear seven helix bundle friends, The TM7 server at EMBL got updated. The multiple sequence alignments now contain many more than 700 sequences. Several sub-classes of GPCRs have been aligned separately. All alignments got one numbering scheme. The sequences of about 450 GPCRs were extracted from SWISS-PROT, and are available from the TM7 server. The sequence profiles used to search the databases and to make the alignments are only available from the FTP site. A tentative alignment of a couple hundred easy to find G-alpha proteins was added, as well as an alignment of 12 bacteriorhodopsin like sequences. If you want a special class of GPCRs to be added, feel free to ask me, it is only a couple hours work to make one.... It would be nice if some of the experts out there would look at the alignments, and tell me where our programs went wrong. Please send comments to "VRIEND@EMBL-Heidelberg.DE", rather than to the server, because the server is just a mail reading program that can only read the syntax as described below. I will later send a summary of the corrections around. About 200 models of the seven helix area of GPCRs remained unaltered. These models are of poor quality, but they are administratively correct, and could serve well as start of your own modeling, or are nice for quickly looking at a model. One of these days I will make 'better' models. In the first year of this service we got 8109 request, and we managed to send the requested file back in almost 90% of all cases. We have problems with mail that comes in via fire-walls, so if your site is fire-wall protected, you better use FTP instead of the mail server. Also, about 15% of all requests does not have the correct format, and can not be dealt with (dont put escape character at the end of server request lines; try not to send your signature because we dont know what to do with those; the requests for tuna fish, fat angus drives, and a few more in this category also were not taken care of adequately by the server...). Please do not send more than 10 requests per day. Please do not request more than one model per request. If you want them all, use FTP. Please be patient, and complain to me only after you did not get a reply within 48 hours. The rest of this mail consists of a legal blurr that unfortunately is required with services like these and some notes about how to get the data. I suggest that those of you who are new users of this mail service send the command: GET HELP:GENERAL.HELP to "TM7@EMBL-Heidelberg.DE" Copyright (C) 1993, 1994, 1995 G. Vriend The TM7 program and data suit is a free data distribution system. It is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY without even the implied warranty of merchantability or fitness for a particular purpose. The copyright holders and/or other parties provide the program "AS IS", without warranty of any kind, either expressed or implied. The entire risk as to the quality and performance of the service is with you. Should any program or data prove defective, you assume the cost of all necessary servicing, repair or correction. The file server holds data related to G-protein coupled receptors that possess the seven helix transmembrane bundle motif. One can freely extract the data. It is allowed to modify and redistribute the data at a limited scale, provided the copyright notices are not removed. (All modifications must be documented and the modifier must be identified and take scientific responsibility for the modification). The data can freely be used, but it is neither allowed to sell the data, nor is it allowed to redistribute them directly or indirectly for commercial purposes. Data and other files that are not produced by G.Vriend (as can be recognized from the "Copyright (C) 1993, 1994, 1995 G. Vriend" notice) can only be extracted for personal use. If you forget to acknowledge me, fine, I dont care, but please give proper acknowledgments to the people that help me maintain this server. From owner-7tms_r@net.bio.net Thu Feb 16 22:00:00 1995 Path: biosci!sb.com!Jonathan_A_Lee%notes From: Jonathan_A_Lee%notes@sb.com (Jonathan A Lee) Newsgroups: bionet.molbio.proteins.7tms_r Subject: CALCITONIN RECEPTORS Date: 17 Feb 1995 10:26:50 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 22 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502172125.AA1389@pho018.sb.com> NNTP-Posting-Host: net.bio.net I would like to identify individuals that can help me obtain cDNAs encoding human calcitonin receptor (either or both splice variants) and stable cell lines expressing the same. In addition, is there any new information concerning the splice variants of calcitonin receptors from any species? Thanks. Jonathan A. Lee, Ph.D. Department of Macromolecular Science Mail Code: UE0447d SmithKline Phamaceuticals 709 Swedeland Road King of Prussia, PA 19406 USA phone: 610-270-7588 FAX: 610-270-4091 E Mail: LEEJA%PHVAX.DNET@SB.COM From owner-7tms_r@net.bio.net Sat Feb 18 22:00:00 1995 Path: biosci!agate!dog.ee.lbl.gov!overload.lbl.gov!ames!waikato!comp.vuw.ac.nz!waltec.warc.cri.nz!fidlera From: fidlera@waltec.warc.cri.nz (Andrew Fidler) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Stem Cell Factor Antibodies Date: Mon, 20 Feb 1995 15:50:02 Organization: AgResearch Wallaceville Lines: 4 Message-ID: NNTP-Posting-Host: pc155.warc.cri.nz Summary: Antibody search Keywords: Steel Stem X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Does anyone out there know of a source of antibodies against a mammalian stem cell factor (SCF) (i.e. murine "steel") Regards, Andrew Fidler From owner-7tms_r@net.bio.net Sun Feb 19 22:00:00 1995 Path: biosci!WELCHLINK.WELCH.JHU.EDU!mlevine From: mlevine@WELCHLINK.WELCH.JHU.EDU (MICHAEL A LEVINE) Newsgroups: bionet.molbio.proteins.7tms_r Subject: adenylyl cyclase Date: 20 Feb 1995 22:24:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Does anyone out there know of decent antibodies that can detect different forms of mammalian adenylyl cyclases that are normally expressed in plasma membranes? Michael Levine, M.D. Johns Hopkins/ Endocrine mlevine@welchlink.welch.jhu.edu 410-955-7233 (voice) 410-955-0841 (fax) From owner-7tms_r@net.bio.net Mon Feb 20 22:00:00 1995 Path: biosci!GSBS.GS.UTH.TMC.EDU!dclark From: dclark@GSBS.GS.UTH.TMC.EDU (Dick Clark) Newsgroups: bionet.molbio.proteins.7tms_r Subject: partial agonists Date: 21 Feb 1995 16:44:55 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502220044.AA15650@gsbs> NNTP-Posting-Host: net.bio.net RE Steven Seiler's questions: Our group (Whaley et al Mol Pharm 45:481-489) has developed a model that quite nicely describes beta receptor activation of AC over a 1000 fold range of R levels. In this model the rate of activation of AC is equal to kon x R* (where kon is the rate constant for activation of AC and R* is the active conformer of R). In the context of this model the definition of a partial agonist is the ratio of its kon x R* to that of a full agonist. It should be noted that some pharmacologists still attempt to define partial agonism by efficacy (Vmax) alone. This is incorrect, since we and others have shown that a partial agonist looks like a full agonist if R levels are sufficiently high. There is not yet resolution of whether partial agonists are simply not as good at stabilizing the R* activated state of the receptor (the Monod model), or whether they actually induce different, less efficient conformations of R (the Koshland model). We favor the Monod model as do others in the field (eg. see Samama et al JBC 268:4625). For a more theoretical treatment of the differences between partial and full agonists with regard to the important concept of "encounter time" see Stickle and Barber Mol Pharm 40:276-288. With regard to the use of GTPase activity for the measurement of partial agonism: this is of course a measure of koff, not kon, and there are the notorious difficulties in measuring hormone/receptor activated GTPase in membranes. The GTPase assays almost invariably requires pure components. Dick Clark From owner-7tms_r@net.bio.net Mon Feb 20 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!ns1.faseb.org!darwin.sura.net!gatekeeper.es.dupont.com!eplrx7!eplrx7!gonyege From: gonyege@lldmpc.dnet.dupont.com (Gregory E. Gonye PhD) Subject: A endocytosis poison for use in "live" binding assay Message-ID: Followup-To: gonyege@lldmpc.dnet.dupont.com Sender: news@eplrx7.es.duPont.com (News Admin Account) Nntp-Posting-Host: gonye.es.dupont.com Organization: DMPC X-Newsreader: VersaTerm Link v1.1.1 Date: Tue, 21 Feb 1995 22:43:53 GMT Lines: 8 Hello everyone, I am attempting to develop a radioligand binding assay on "live" tissue. One major hurdle is endocytosis. If the assay is done at 0C the binding kinetics will be too slow, not to mention the state change in the membranes, but if I run the assay at higher temps endocytosis will make the data difficult to interpret. Therefore I am searching for an endocytosis poison with good penetrance in order to block endocytosis at RT. Please help. Thanks. G. From owner-7tms_r@net.bio.net Tue Feb 21 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: Cornelius Krasel Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re; endocytosis poison Date: 22 Feb 1995 22:29:12 -0000 Lines: 16 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3igdro$qpt@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Gregory E. Gonye (gonyege@llcmpc.dnet.dupont.com) asked for inhibitors of endocytosis. Several chemicals have been reported to block endocytosis of G-protein-coupled receptors. Among those are concanavalin A, high sucrose in the medium and agents that deplete the cells of endogenous ATP (like phenylarsinoxide). A really specific poison is not available, however; endocytosis of GPCRs is quite an unresolved issue nowadays. Hope that helps, --Cornelius. -- /* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */ /* email: krasel@alf.biochem.mpg.de fax: +49 89 8578 3795 */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Tue Feb 21 22:00:00 1995 Path: biosci!NEURO.MSKCC.ORG!brown From: brown@NEURO.MSKCC.ORG Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 22 Feb 1995 18:31:20 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <199502230231.SAA24933@net.bio.net> NNTP-Posting-Host: net.bio.net help