From owner-7tms_r@net.bio.net Tue Aug 01 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!news.sprintlink.net!newsfeed.internetmci.com!uunet!in1.uu.net!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: biobone@aol.com (Bio Bone) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Antibodies to 7TMS Date: 2 Aug 1995 18:01:14 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 12 Sender: root@newsbf02.news.aol.com Message-ID: <3vosja$j09@newsbf02.news.aol.com> Reply-To: biobone@aol.com (Bio Bone) NNTP-Posting-Host: newsbf02.mail.aol.com I am looking for a number of antibodies (either monoclonal or polyclonal) to the following receptors. (I have looked in Linscotts Dir. and a number of catalogs): Histamine Receptor Serotonin Receptor alpha adrenergic receptor Platelet Activating Factor Receptor. Thanks Bradley From owner-7tms_r@net.bio.net Wed Aug 02 23:00:00 1995 Path: biosci!bcm!news.msfc.nasa.gov!newsfeed.internetmci.com!news.sprintlink.net!nwfocus1.wa.com!news.halcyon.com!usenet From: Gary McKnight Newsgroups: bionet.molbio.proteins.7tms_r Subject: Wanted: G alpha-S ligand/R list Date: 3 Aug 1995 16:46:45 GMT Organization: ZymoGenetics Lines: 13 Message-ID: <3vquhl$1re@news.halcyon.com> NNTP-Posting-Host: garym.zgi.com 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.7tms_r I am need of a list of G-protein coupled receptors and/or ligands which couple through G alpha S. If you can help provide such answers please contact me via email. Thanks in advance for all of your help. Gary McKnight ZymoGenetics garym@zgi.com From owner-7tms_r@net.bio.net Wed Aug 02 23:00:00 1995 Path: biosci!DOLPHIN.UPENN.EDU!stein From: stein@DOLPHIN.UPENN.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: postdoc position-wanted Date: 3 Aug 1995 17:20:27 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 37 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508040018.UAA27018@dolphin.upenn.edu> NNTP-Posting-Host: net.bio.net From stein Thu Aug 3 19:54:23 1995 >From stein@dolphin.upenn.edu Thu Aug 3 19:54:23 1995 Received: by dolphin.upenn.edu id TAA25423; Thu, 3 Aug 1995 19:54:23 -0400 Posted-Date: Thu, 3 Aug 1995 19:54:23 -0400 Received-Date: Thu, 3 Aug 1995 19:54:23 -0400 Subject: tr To: stein@dolphin.upenn.edu Date: Thu, 3 Aug 1995 19:54:23 -0400 (EDT) From: stein@alumni.upenn.edu X-Sender: "stein" X-Mailer: ELM [version 2.4 PL23-upenn2.9] MIME-Version: 1.0 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Length: 430 Status: RO *** Please, don't response by reply command *** SUBJECT: Seeking for postdoctoral position in US FIELD: Molecular Biology & Genetics of Oncogenes or Antioncogenes SKILLS: Cloning, sequencing, mutagenesis, PCR (RT-PCR), Southern & Northern blotting, immunoprecipitation, CAT, tissue culture, tumor induction, PC, publications registered in CC, etc. WHEN: available immediately, further information upon request ADDRESS: stein@dolphin.upenn.edu From owner-7tms_r@net.bio.net Wed Aug 02 23:00:00 1995 Path: biosci!GIBBS.OIT.UNC.EDU!laiter From: laiter@GIBBS.OIT.UNC.EDU (Sergei Laiter) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Pro kinks in helices Date: 3 Aug 1995 10:56:36 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear Netters, I'm modeling GPCR's and having a question at the moment. In a few relatively recent publications I've read about introducing pro kinks in the helices (e.g. setting the main chain torsions for the residue before a pro to -38 and -64. I understand that this is based on observations from structures with good resolution. However, when I checked for such kinks in the BACTERIORHODOPSIN, I did not see this kinks. Could it be because of the low resolution or maybe helices inside of a membrane are different in this respect? Any comments on this would be appreciated. -Sergei From owner-7tms_r@net.bio.net Wed Aug 02 23:00:00 1995 Path: biosci!aol.com!lldmpc::mrgate::a1::kellerma%lldmpc.dnet.dupont.com From: lldmpc::mrgate::a1::kellerma%lldmpc.dnet.dupont.com@aol.com Newsgroups: bionet.molbio.proteins.7tms_r Subject: unsubscribe Date: 3 Aug 1995 10:56:16 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: <950803135009_129641880@aol.com> NNTP-Posting-Host: net.bio.net From: NAME: MARGARET A. KELLER FUNC: TEL: To: "7tms_r@net.bio.net@aol.com"@esds01@mrgate unsubscribe From owner-7tms_r@net.bio.net Thu Aug 03 23:00:00 1995 Path: biosci!bath.ac.uk!bsscw From: bsscw@bath.ac.uk (C Wood) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Pro kinks in helices Date: 4 Aug 1995 05:01:52 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 51 Sender: daemon@net.bio.net Distribution: world Message-ID: References: NNTP-Posting-Host: net.bio.net Sergei - be very wary of using bR to model kinks - the structure is full of steric clashes - if you use the back-bone co-ordinates of bR you will have a hell of a time energy minimising it ... i helped someone out who did just this and we rebuilt his transmembrane helices and he finally realised that the back-bone co-ordinates of bR - especially the ones with middle prolines are pretty hopeless as a template for modellin putative GPCR transmembrane helices ... get in touch with me by email ... better still ... order my Ph.D. thesis from Glasgow University Library (or maybe the British Library) ... you can also have complete access to my transmembrane models if you want them ... there are some very good papers that can help you in your modelling too ... email me ___________________________________________________________________________ To preserve the environment this message was ___ _ _ _ _ _ | _ \_ _(_)_ _| |_ ___ __| | ___ _ _ _ _ ___ __ _ _ __| |___ __| | | _/ '_| | ' \ _/ -_) _` | / _ \ ' \ | '_/ -_) _| || / _| / -_) _` | |_| |_| |_|_||_\__\___\__,_| \___/_||_| |_| \___\__|\_, \__|_\___\__,_| |__/ _ _ ___| |___ __| |_ _ _ ___ _ _ ___ Chris Wood PhD / -_) / -_) _| _| '_/ _ \ ' \(_-< Email: bsscw@bath.ac.uk \___|_\___\__|\__|_| \___/_||_/__/ Biochem Dept., Bath Uni., Bath, UK, BA2 7AY. Tel work: UK 01225-826826 ext 5118 ___________________________________________________________________________ On 3 Aug 1995, Sergei Laiter wrote: > > Dear Netters, > > I'm modeling GPCR's and having a question at the moment. > In a few relatively recent publications I've read about introducing pro > kinks in the helices (e.g. setting the main chain torsions for the > residue before a pro to -38 and -64. I understand that this is based on > observations from structures with good resolution. However, when I > checked for such kinks in the BACTERIORHODOPSIN, I did not see this kinks. > Could it be because of the low resolution or maybe helices inside of a > membrane are different in this respect? > > Any comments on this would be appreciated. > > -Sergei > From owner-7tms_r@net.bio.net Thu Aug 03 23:00:00 1995 Path: biosci!VAX.BIO.LEEDS.AC.UK!BMB6HDD From: BMB6HDD@VAX.BIO.LEEDS.AC.UK Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 4 Aug 1995 07:10:54 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <23402932_000E0A10.009945EDF982E080$7_1@UK.AC.LEEDS.BIO.VAX> NNTP-Posting-Host: net.bio.net I'm probably preaching to the converted but can I recommend the review by Terry Kenakin in TiPS 16:232-238. It summarises much of the latest views on GPCR states etc. Dan Donnelly Dept. Pharmacology University of Leeds Leeds LS2 9JT UK d.donnelly@leeds.ac.uk From owner-7tms_r@net.bio.net Thu Aug 03 23:00:00 1995 Path: biosci!UMDNJ.EDU!reza From: reza@UMDNJ.EDU (Syed Reza) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 4 Aug 1995 13:30:35 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508042030.NAA29254@net.bio.net> NNTP-Posting-Host: net.bio.net subscribe From owner-7tms_r@net.bio.net Thu Aug 03 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!newsfeed.internetmci.com!solaris.cc.vt.edu!news.duke.edu!godot.cc.duq.edu!newsfeed.pitt.edu!dsinc!news.ee.vill.edu!news From: Brian Shoop Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ligand Software for Windows? - Is there any Date: 4 Aug 1995 15:56:29 GMT Organization: Weis Center for Research Lines: 10 Message-ID: <3vtfvd$jjr@ftp.ee.vill.edu> NNTP-Posting-Host: bshoop.geisinger.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) I have heard that there is a Windows version of the ligand program for receptor binding analysis. Has anyone heard of it? Please reply via email. - Brian Shoop Weis Center for Research Danville, PA 17822-2600 bshoop@smtp.geisinger.edu From owner-7tms_r@net.bio.net Thu Aug 03 23:00:00 1995 Path: biosci!EMBL-HEIDELBERG.DE!Gert.Vriend From: Gert.Vriend@EMBL-HEIDELBERG.DE Newsgroups: bionet.molbio.proteins.7tms_r Subject: States Date: 4 Aug 1995 09:29:25 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HTOO0J268KIHI54T@EMBL-Heidelberg.DE> NNTP-Posting-Host: net.bio.net We postulated a much simpeler (probably too simple, but lacking data, the simpler the better I would say) model for "the active and inactive" state of GPCRs. See TIPS, june 1994, Vol 15, 170-172 By Oliveira et al. Comments on this article are welcome! Gert Vriend. From owner-7tms_r@net.bio.net Mon Aug 07 23:00:00 1995 Path: biosci!ic.ac.uk!p.little From: p.little@ic.ac.uk (P.Little) Newsgroups: bionet.molbio.proteins.7tms_r Subject: job in embryology and GCRs Date: 8 Aug 1995 03:11:59 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <9508081009.AA20596@bccmsa.bc.ic.ac.uk> NNTP-Posting-Host: net.bio.net Position available Molecular biology of neural development - involvment of a new GCR. Post doctoral position to work on a family of newly discovered 7-pass transmembrane G-protein coupled receptors in the mouse. One receptor is expressed prior to gastrulation and its expression is tightly controlled in the developing CNS. The project is to characterise the ligand of the receptor, its function and to establish the limits of the family. Work on the human and fruit fly homologue will also feature. Experience in any or all of the following techniques would be an advantage: in situ hybridisation, analysis of mouse development and molecular biology techniques. Inquires to Dr. Peter Little, tel. 0171 594 5288, fax. 0171 823 7525, E-mail p.little@ic.ac.uk. No application forms: send CV plus names and phone numbers of two referees to Dr. Little at Department of Biochemistry, Imperial College, London SW7 2AZ, UK. From owner-7tms_r@net.bio.net Tue Aug 08 23:00:00 1995 Path: biosci!CODON.NIH.GOV!usdin From: usdin@CODON.NIH.GOV (Ted Usdin) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoctoral position available Date: 9 Aug 1995 05:59:39 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 39 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Postdoctoral position I have a position available within my small group in the Laboratory of Cell Biology at NIMH. This is a large laboratory with a variety of available techniques and ample opportunities for collaboration. We have recently cloned several members of the secretin receptor family, including the GIP receptor, VIP2 receptor and PTH2 receptor. We now plan on using these and other members of the secretin receptor family to study ligand interaction and signal transduction within this receptor family. We also hope to learn more about the physiological roles of these receptors, particularly in the brain. This is an opportunity for someone capable of independent work who has a strong background in pharmacology, molecular biology, or neuroscience. I would appreciate receiving a c.v. and names of references from anyone interested, either by e-mail or conventional mail. Thank you for bringing this to the attention of any students who you think may be good candidates. Yours truly, Ted Usdin ****************************** Ted B. Usdin, M.D., Ph.D. Laboratory of Cell Biology, NIMH Bldg 36/Rm3A17 {36 Convent Dr MSC 4090 (U.S. mail)} BETHESDA MD 20892-4090 e-mail usdin@codon.nih.gov tel 301-402-4161 fax 301-402-1748 From owner-7tms_r@net.bio.net Sun Aug 13 23:00:00 1995 Path: biosci!RECEPTOR.MGH.HARVARD.EDU!lfk From: lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: JBC TOC 8/18-9/1 Excerpted for 7tms_r Date: 14 Aug 1995 05:27:36 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 108 Sender: daemon@net.bio.net Distribution: world Message-ID: <9508141217.AA15036@receptor> NNTP-Posting-Host: net.bio.net AU Lopez-I. Burns-D-J. Lambeth-J-D. TI Regulation of phospholipase D by protein kinase C in human neutrophils. Conventional isoforms of protein kinase C phosphorylate a phospholipase D-related component in the plasma membrane. SO J-Biol-Chem. 1995 August 18. 270(33). p 19465. AU Power-C-A. Meyer-A. Nemeth-K. Bacon-K-B. Hoogewerf-A-J. Proudfoot-A-E-I. Wells-T-N-C. TI Molecular cloning and functional expression of a novel CC chemokine receptor cDNA from a human basophilic cell line SO J-Biol-Chem. 1995 August 18. 270(33). p 19495. AU Liu-J. Schoneberg-T. Rhee-M-. Wess-J. TI Mutational analysis of the relative orientation of transmembrane helices I and VII in G protein-coupled receptors SO J-Biol-Chem. 1995 August 18. 270(33). p 19532. AU Buhl-A-M. Osawa-S. Johnson-G-L. TI Mitogen-activated protein kinase activation requires two signal inputs from the human anaphylatoxin C5a receptor SO J-Biol-Chem. 1995 August 25. 270(34). p 19828. AU Hershenson-M-B. Chao-T-S-O. Abe-M-K. Gomes-I. Kelleher-M-D. Solway-J. Rosner-M-R. TI Histamine antagonizes serotonin and growth factor-induced mitogen-activated protein kinase activation in bovine tracheal smooth muscle cells SO J-Biol-Chem. 1995 August 25. 270(34). p 19908. AU Schmidt-M. Fasselt-B. Rumenapp-U. Bienek-C. Wieland-T. Koppen-C-J-. Jakobs-K-H. TI Rapid and persistent desensitization of m3 muscarinic acetylcholine receptor-stimulated phospholipase D. Concomitant sensitization of phospholipase C SO J-Biol-Chem. 1995 August 25. 270(34). p 19949. AU Ptasznik-A. Traynor-Kaplan-A. Bokoch-G-M. TI G protein-coupled chemoattractant receptors regulate Lyn tyrosine kinaseShc adapter protein signaling complexes SO J-Biol-Chem. 1995 August 25. 270(34). p 19969. AU Pond-L. Kuhn-L-A. Teyton-L. Schutze-M-P. Tainer-J-A. Jackson-M-R. Peterson-P-A. TI A role for acidic residues in di-leucine motif-based targeting to the endocytic pathway SO J-Biol-Chem. 1995 August 25. 270(34). p 19989. AU Azarani-A. Goltzman-D. Orlowski-J. TI Parathyroid hormone and parathyroid hormone-related peptide inhibit the apical Na+/H+ exchanger NHE-3 isoform in renal cells (OK) via a dual signaling cascade involving protein kinase A and C SO J-Biol-Chem. 1995 August 25. 270(34). p 20004. AU Cosowsky-L. Rao-S-N-V. Macdonald-G-J. Papkoff-H. Campbell-R-K. Moyle-W-R. TI The groove between the {alpha}- and {beta}-subunits of hormones with lutropin (LH) activity appears to contact the LH receptor, and its conformation is changed during hormone binding SO J-Biol-Chem. 1995 August 25. 270(34). p 20011. AU Moyle-W-R. Campbell-R-K. Rao-S-N-V. Ayad-N-G. Bernard-M-P. Han-Y. Wang-Y. TI Model of human chorionic gonadotropin and lutropin receptor interaction that explains signal transduction of the glycoprotein hormones SO J-Biol-Chem. 1995 August 25. 270(34). p 20020. AU Murasawa-S. Matsubara-H. Kijima-K. Maruyama-K. Mori-Y. Inada-M. TI Structure of the rat V1a vasopressin receptor gene and characterization of its promoter region and complete cDNA sequence of the 3{prime}-end SO J-Biol-Chem. 1995 August 25. 270(34). p 20042. AU Aragay-A-M. Collins-L-R. Post-G-R. Watson-A-J. Feramisco-J-R. Brown-J-H. Simon-M-I. TI G12 requirement for thrombin-stimulated gene expression and DNA synthesis in 1321N1 astrocytoma cells SO J-Biol-Chem. 1995 August 25. 270(34). p 20073. AU Horiuchi-M. Koike-G. Yamada-T. Mukoyama-M. Nakajima-M. Dzau-V-J. TI The growth-dependent expression of angiotensin II type 2 receptor is regulated by transcription factors interferon regulatory factor-1 and -2 SO J-Biol-Chem. 1995 August 25. 270(34). p 20225. AU Kohout-T-A. Rogers-T-B. TI Angiotensin II activates the Na+/HCO3- symport through a phosphoinositide-independent mechanism in cardiac cells SO J-Biol-Chem. 1995 September 1. 270(35). p 20432. AU Rivkees-S-A. Lasbury-M-E. Barbhaiya-H. TI Identification of domains of the human A1 adenosine receptor that are important for binding receptor subtype-selective ligands using chimeric A1/A2a adenosine receptors SO J-Biol-Chem. 1995 September 1. 270(35). p 20485. AU Herzig-M-C-S. Leeb-Lundberg-L-M-F. TI The agonist binding site on the bovine bradykinin B2 receptor is adjacent to a sulfhydryl and is differentiated from the antagonist binding site by chemical cross-linking SO J-Biol-Chem. 1995 September 1. 270(35). p 20591. AU McKinnon-L-A. Nathanson-N-M. TI Tissue-specific regulation of muscarinic acetylcholine receptor expression during embryonic development SO J-Biol-Chem. 1995 September 1. 270(35). p 20636. From owner-7tms_r@net.bio.net Sun Aug 13 23:00:00 1995 Path: biosci!CELLBIO.DUKE.EDU!stephen_ferguson From: stephen_ferguson@CELLBIO.DUKE.EDU ("Stephen Ferguson") Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 14 Aug 1995 11:49:29 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Stephen Ferguson Fergu007@mc.duke.edu subscribe From owner-7tms_r@net.bio.net Sun Aug 13 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Mitochondrial receptors Date: 14 Aug 1995 10:21:09 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 33 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508141718.NAA21141@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > > I originally started (or tried to) start the discussion on intracellular 7TMD > receptors on the 7TMD receptor newsgroup. I just wanted to ask you directly > just what did you mean by "Setting aside the question of mitochondrial > receptors..". Are there well-characterized mitochondrial 7TMD receptors? > Thanks for you help. > Roberto. > > P.S. I'm tracing those ref.s you posted. They seem interesting! Mitochondria do not express any 7tms_r to my knowledge. Some examples of apparent receptors present on mitochondria are the peripheral benzodiaepine receptor, which particicpates in the control of steroid biosynthesis in the adrenal cortex (many of the references for this are from Sol Snyder's group). Receptors are often present in "mitochondrial fractions" but this is probably due to contamination with golgi and plasma membrane, which have similar densities to mitochondria. 7tms_r are sometimes seen on mitochondria with electron immunocytomchemistry, but this is usually felt to be an artifact of the fixation process, whereby a fragment of nearby membrane might become cross-linked to mitochondria during aldehyde fixation. -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* From owner-7tms_r@net.bio.net Sun Aug 13 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!daresbury!nntp-trd.UNINETT.no!Norway.EU.net!EU.net!howland.reston.ans.net!tank.news.pipex.net!pipex!warwick!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (AJ. Collier) Subject: Pertussis sensitive G-proteins weights Message-ID: Sender: usenet@uns.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, 14 Aug 1995 15:58:46 GMT Lines: 6 I have been looking at inhibitory G-proteins in keratinocyte cell lines by [32p] ADP ribosylation and western blotting. In both cases I get two bands between 40-45kDa. What are the molecular masses of G-proteins shown to be sensitive to pertussis toxin. Thanks From owner-7tms_r@net.bio.net Mon Aug 14 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!news.sprintlink.net!EU.net!Germany.EU.net!nntp.gmd.de!newsserver.jvnc.net!netnews.sb.com!news From: Steven_Mcclue-1%notes@sb.com (Steve Mcclue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Pertussis sensitive G-proteins weights Date: 15 Aug 1995 07:54:01 GMT Organization: Smithkline Beecham Pharmaceuticals Lines: 16 Message-ID: <40pjqp$27f@phunn1.sb.com> References: NNTP-Posting-Host: had542.uk.sb.com Mime-Version: 1.0 X-Newsreader: WinVN 0.99.2 In article , dhajc@zeus.bris.ac.uk says... > >I have been looking at inhibitory G-proteins in keratinocyte cell lines >by [32p] ADP ribosylation and western blotting. In both cases I get two >bands between 40-45kDa. What are the molecular masses of G-proteins shown >to be sensitive to pertussis toxin. >Thanks > Gi1, Gi2 and Gi3 fall into the MW range you have suggested (Go is 39kDa, so may be outside your range if you're sure about the 40kDa cutoff). Gi1 and Gi3 are usually detected at about 41kDa, while Gi2 comes in at about 40kDa. If you really want to know what you've got, you'll need to get hold of some anti-G protein antibodies. Steve From owner-7tms_r@net.bio.net Mon Aug 14 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: ptaylor@hgmp.mrc.ac.uk (Dr. P.L. Taylor) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoc position - protein purification / receptor biochemistry Date: 15 Aug 1995 17:25:23 +0100 Lines: 22 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <40qhpj$1b5@mserv1.dl.ac.uk> content-length: 1186 Original-To: 7tms_r@dl.ac.uk NON-CLINICAL SCIENTIST GRADE II Applications are invited for the above position to join a team investigating the structure and function of pituitary releasing-hormone receptors relevant to reproduction. The work will involve the over-expression of cloned receptors, isolation and purification of receptor proteins with a view to eventually solving structures by crystallographic means. PhD in a relevant subject, and experience of protein purification are essential. This is a three year appointment, salary range (Pounds sterling) 16628 - 19848. The position is available immediately, applications close August 21, 1995. The MRC is an equal-opportunity employer. Applications may be addressed by EMail to business.manager@ed-rbu.mrc.ac.uk, or mailed to Patricia Young at the address below. For further information, EMail me. Phil Taylor | MRC Reproductive Biology Unit | Centre for Reproductive Biology | 37 Chalmers Street mbplt@seqnet.dl.ac.uk | Edinburgh EH3 9EW ptaylor@hgmp.mrc.ac.uk | Scotland. From owner-7tms_r@net.bio.net Mon Aug 14 23:00:00 1995 Path: biosci!FTN.NET!kirbyr From: kirbyr@FTN.NET (Robert Kirby) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Subscribe Date: 15 Aug 1995 12:44:19 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 6 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Please subscribe me to the TM7 group. Robert Kirby my e-mail is kirbyr@ftn.net thanks From owner-7tms_r@net.bio.net Mon Aug 14 23:00:00 1995 Path: biosci!UCDAVIS.EDU!jhchen From: jhchen@UCDAVIS.EDU (Jin Chen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor Subtypes Date: 15 Aug 1995 23:26:34 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508160624.XAA19285@peseta.ucdavis.edu> NNTP-Posting-Host: net.bio.net Can anyone tell me the criteria of defining receptor subtypes (particularly GCRs)? Thanks Jin From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!bcm!news.msfc.nasa.gov!newsfeed.internetmci.com!news.uoregon.edu!tank.news.pipex.net!pipex!howland.reston.ans.net!swrinde!emory!usenet From: Xiaofei Wang Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor Subtypes Date: 16 Aug 1995 17:44:16 GMT Organization: Biomolecular Computing Resource, Emory University Lines: 9 Message-ID: <40tapg$cmc@emory.mathcs.emory.edu> References: <199508160624.XAA19285@peseta.ucdavis.edu> NNTP-Posting-Host: bimcore.emory.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (X11; I; SunOS 5.4 sun4m) X-URL: news:199508160624.XAA19285@peseta.ucdavis.edu Hi there: From what I know, receptors can be classified according to their pharmacological profiles, i.e., the Kd value to various agonists and antagonists. This criterie is very suitable to G-protein coupled receptors. And for GCRs, I remember they can be classified into 7 groups starting from bacteriorhodopsin, including beta-ARs etc. Another criterie people use is sequence analysis. Just think about how alpha-ARs subtypes are named in alpha-1A, B & D. From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Synaptic Pharmaceutical Co. Date: 16 Aug 1995 08:23:49 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 30 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508161521.LAA05003@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > Just wondering if other people have had problems with Synaptic. I requested a sample of their new (published) alpha-1a selective antagonist SNAP 5089. I just got a call from Dr. Charles Gluchowski who said that their collaboration with Merck won't allow them to release the compound for at least six months more. > > This is fairly unprecedented in my experience, as companies are usually eager to get information on compounds they are interested in, once appropriate attribution and patents are filed. > > Anyone care to comment? > Ken- Synaptic is not a major pharmaceutical house and does not have the wherewithal for complete drug development. They live by technology transfer, which undoubtedly makes things difficult for them. I have worked with these folks and I think they are on the level. I would take the issue up with Merck. Delays in availability up to year are fairly typical, in my experience. Often a company will agree to provide a compound, then take up to a year to "process the request". -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!swrinde!emory!usenet From: kminnem@bimcore.emory.edu (Kenneth Minneman) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Synaptic Pharmaceutical Co. Date: 16 Aug 1995 12:40:12 GMT Organization: Biomolecular Computing Resource, Emory University Lines: 7 Distribution: world Message-ID: <40sovc$b8@emory.mathcs.emory.edu> Reply-To: kminnem@bimcore.emory.edu NNTP-Posting-Host: bimcore.emory.edu Just wondering if other people have had problems with Synaptic. I requested a sample of their new (published) alpha-1a selective antagonist SNAP 5089. I just got a call from Dr. Charles Gluchowski who said that their collaboration with Merck won't allow them to release the compound for at least six months more. This is fairly unprecedented in my experience, as companies are usually eager to get information on compounds they are interested in, once appropriate attribution and patents are filed. Anyone care to comment? From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!bath.ac.uk!bsscw From: bsscw@bath.ac.uk (C Wood) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: chi values of sidechains Date: 16 Aug 1995 23:32:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 85 Sender: daemon@net.bio.net Distribution: world Message-ID: References: <9508162327.AA20900@unlinfo.unl.edu> NNTP-Posting-Host: net.bio.net If any of you guys are using this paper to guide your modelling of side-chain torsion angles: TI: KNOWLEDGE BASED MODELING OF HOMOLOGOUS PROTEINS .2. RULES FOR THE CONFORMATIONS OF SUBSTITUTED SIDECHAINS AU: SUTCLIFFE_MJ, HAYES_FRF, BLUNDELL_TL NA: UNIV LONDON BIRKBECK COLL,DEPT CRYSTALLOG,MOLEC BIOL LAB,MALET ST,LONDON WC1E 7HX,ENGLAND JN: PROTEIN ENGINEERING, 1987, Vol.1, No.5, pp.385-392 For modeling side-chain torsion angles (in alpha-helices - i will post the ones for beta-sheet later) ... read on! [obviously you will have to remove bad contacts and energy minimising etc. etc.] ... i can do this for a small fee ... *laughs* For users of Insight I (possibly Insight II) ... then this command file might save you a lot of time and help prevent marriage breakdown ... :) !! **************************cut here******************************************* set dihedral -69.00 ala_b0:arg*:n ala_b0:arg*:ca ala_b0:arg*:cb ala_b0:arg*:cg set dihedral 175.00 ala_b0:arg*:ca ala_b0:arg*:cb ala_b0:arg*:cg ala_b0:arg*:cd set dihedral 174.00 ala_b0:arg*:cb ala_b0:arg*:cg ala_b0:arg*:cd ala_b0:arg*:ne set dihedral -75.00 ala_b0:arg*:cg ala_b0:arg*:cd ala_b0:arg*:ne ala_b0:arg*:cz set dihedral 177.00 ala_b0:arg*:cd ala_b0:arg*:ne ala_b0:arg*:cz ala_b0:arg*:nh1 set dihedral -70.00 ala_b0:asn*:n ala_b0:asn*:ca ala_b0:asn*:cb ala_b0:asn*:cg set dihedral -49.00 ala_b0:asn*:ca ala_b0:asn*:cb ala_b0:asn*:cg ala_b0:asn*:od1 set dihedral -69.00 ala_b0:asp*:n ala_b0:asp*:ca ala_b0:asp*:cb ala_b0:asp*:cg set dihedral -3.00 ala_b0:asp*:ca ala_b0:asp*:cb ala_b0:asp*:cg ala_b0:asp*:od1 set dihedral 176.00 ala_b0:cys*:n ala_b0:cys*:ca ala_b0:cys*:cb ala_b0:cys*:sg set dihedral -72.00 ala_b0:gln*:n ala_b0:gln*:ca ala_b0:gln*:cb ala_b0:gln*:cg set dihedral 174.00 ala_b0:gln*:ca ala_b0:gln*:cb ala_b0:gln*:cg ala_b0:gln*:cd set dihedral 2.00 ala_b0:gln*:cb ala_b0:gln*:cg ala_b0:gln*:cd ala_b0:gln*:oe1 set dihedral -172.00 ala_b0:glu*:n ala_b0:glu*:ca ala_b0:glu*:cb ala_b0:glu*:cg set dihedral -179.00 ala_b0:glu*:ca ala_b0:glu*:cb ala_b0:glu*:cg ala_b0:glu*:cd set dihedral 159.00 ala_b0:glu*:cb ala_b0:glu*:cg ala_b0:glu*:cd ala_b0:glu*:oe1 set dihedral -67.00 ala_b0:his*:n ala_b0:his*:ca ala_b0:his*:cb ala_b0:his*:cg set dihedral 74.00 ala_b0:his*:ca ala_b0:his*:cb ala_b0:his*:cg ala_b0:his*:nd1 set dihedral -60.00 ala_b0:ile*:n ala_b0:ile*:ca ala_b0:ile*:cb ala_b0:ile*:cg1 set dihedral 172.00 ala_b0:ile*:ca ala_b0:ile*:cb ala_b0:ile*:cg1 ala_b0:ile*:cd1 set dihedral -68.00 ala_b0:leu*:n ala_b0:leu*:ca ala_b0:leu*:cb ala_b0:leu*:cg set dihedral 179.00 ala_b0:leu*:ca ala_b0:leu*:cb ala_b0:leu*:cg ala_b0:leu*:cd1 set dihedral -70.00 ala_b0:lys*:n ala_b0:lys*:ca ala_b0:lys*:cb ala_b0:lys*:cg set dihedral -177.00 ala_b0:lys*:ca ala_b0:lys*:cb ala_b0:lys*:cg ala_b0:lys*:cd set dihedral 176.00 ala_b0:lys*:cb ala_b0:lys*:cg ala_b0:lys*:cd ala_b0:lys*:ce set dihedral -177.00 ala_b0:lys*:cg ala_b0:lys*:cd ala_b0:lys*:ce ala_b0:lys*:nz set dihedral -172.00 ala_b0:met*:n ala_b0:met*:ca ala_b0:met*:cb ala_b0:met*:cg set dihedral -177.00 ala_b0:met*:ca ala_b0:met*:cb ala_b0:met*:cg ala_b0:met*:sd set dihedral 67.00 ala_b0:met*:cb ala_b0:met*:cg ala_b0:met*:sd ala_b0:met*:ce set dihedral -64.00 ala_b0:phe*:n ala_b0:phe*:ca ala_b0:phe*:cb ala_b0:phe*:cg set dihedral -91.00 ala_b0:phe*:ca ala_b0:phe*:cb ala_b0:phe*:cg ala_b0:phe*:cd1 set dihedral -65.00 ala_b0:ser*:n ala_b0:ser*:ca ala_b0:ser*:cb ala_b0:ser*:og set dihedral 63.00 ala_b0:thr*:n ala_b0:thr*:ca ala_b0:thr*:cb ala_b0:thr*:og1 set dihedral -160.00 ala_b0:trp*:n ala_b0:trp*:ca ala_b0:trp*:cb ala_b0:trp*:cg set dihedral -104.00 ala_b0:trp*:ca ala_b0:trp*:cb ala_b0:trp*:cg ala_b0:trp*:cd1 set dihedral -67.00 ala_b0:tyr*:n ala_b0:tyr*:ca ala_b0:tyr*:cb ala_b0:tyr*:cg set dihedral -88.00 ala_b0:tyr*:ca ala_b0:tyr*:cb ala_b0:tyr*:cg ala_b0:tyr*:cd1 set dihedral 172.00 ala_b0:val*:n ala_b0:val*:ca ala_b0:val*:cb ala_b0:val*:cg1 ****************************cut here*************************************** this command file will work on object ala_b0 (zero) ..... please edit and apply to any object .... i have only just typed it ... so if anyone spots the obvious error (quite likely!) ... please let me know. ___________________________________________________________________________ To preserve the environment this message was ___ _ _ _ _ _ | _ \_ _(_)_ _| |_ ___ __| | ___ _ _ _ _ ___ __ _ _ __| |___ __| | | _/ '_| | ' \ _/ -_) _` | / _ \ ' \ | '_/ -_) _| || / _| / -_) _` | |_| |_| |_|_||_\__\___\__,_| \___/_||_| |_| \___\__|\_, \__|_\___\__,_| |__/ _ _ ___| |___ __| |_ _ _ ___ _ _ ___ Chris Wood PhD / -_) / -_) _| _| '_/ _ \ ' \(_-< Email: bsscw@bath.ac.uk \___|_\___\__|\__|_| \___/_||_/__/ Biochem Dept., Bath Uni., Bath, UK, BA2 7AY. Tel work: UK 01225-826826 ext 5118 ___________________________________________________________________________ From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!UNLINFO.UNL.EDU!jfrank From: jfrank@UNLINFO.UNL.EDU (julie frank) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Selectable markers for reporter cell line Date: 16 Aug 1995 16:29:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <9508162327.AA20900@unlinfo.unl.edu> NNTP-Posting-Host: net.bio.net I am developing a stable reporter cell line with a recombinant GCR. Has anyone attempted to utilize two selectable markers in a stable cell line such as this one? For example, the receptor plasmid would be selected according to G418 resistance and the reporter gene plasmid would utilize hygromycin as its selectable marker. This would require subcloning the selectable markers into the respective plasmid constructs. From my searches, it appears that most people cotransfect three plasmids in a situation such as this; the reporter plasmid, the receptor plasmid, and pSV2neo. Why is this later approach a better one? Has anyone tried ouabain as a selectable marker? From owner-7tms_r@net.bio.net Tue Aug 15 23:00:00 1995 Path: biosci!UNMCVM.UNMC.EDU!DBYLUND From: DBYLUND@UNMCVM.UNMC.EDU ("David B. Bylund --UNMCVM", DBYLUND) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor subtypes Date: 16 Aug 1995 12:07:24 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 41 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508161907.MAA27993@net.bio.net> NNTP-Posting-Host: net.bio.net FROM: David B. Bylund --UNMCVM(DBYLUND) Jin Chen asked about the criteria for defining receptor subtypes. Historically, the most useful criteria for defining types and subtypes of receptors have been pharmacological, i.e. based on the rank order of potencies of agonists and the relative affinities of antagonists. Other criteria such as anatomical localization (pre- vs post-synaptic) or functional response (excitatory vs inhibitory) have not proven to be useful. More recently, the predicated primary amino acid sequence from cloning and the relative sequence similarities of related receptors have also become useful. My hierarchical method for classifying receptors with examples from the adrenergic receptor family is as follows: Superfamily G-protein coupled; Ligand gated ion channels Family (orClass) Adrenergic Type (or Subfamily) Alpha-1; Alpha-2; Beta Subtype(or member) Beta-1;Beta-2; Beta-3; Alpha-2A; Alpha-2B; Alpha-2C Pharmacological subtypes Alpha-2A and Alpha-2D Each level further down, the lower the degree of selectivity of agonists and antagonist, and the greater the sequence similarity. Although the alpha-2A and alpha-2D have significantly different pharmacological characteristics, they have a high sequence similarity (89%) and only one of them is found in a given species, and thus they are generally not considered to be "true" subtypes. Often, there are correlates to the pharmacological/molecular classification scheme which are not strictly part of the classification criteria. For example, the three adrenergic receptor types each appear to interact preferentially with a distinct family of G-proteins: alpha-1 with Gq; alpha-2 with Gi and beta with Gs. David B. Bylund, Ph.D. Department of Pharmacology University of Nebraska Medical Center P.O. Box 986260 Omaha, NE 68198-6260 voice: 402/559-4788 fax: 402/559-7495 EMAIL: DBYLUND@UNMC.EDU From owner-7tms_r@net.bio.net Thu Aug 17 23:00:00 1995 Path: biosci!VAX.GRC.NIA.NIH.GOV!chahrzad From: chahrzad@VAX.GRC.NIA.NIH.GOV Newsgroups: bionet.molbio.proteins.7tms_r Subject: dominant negative Date: 18 Aug 1995 19:24:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 9 Sender: daemon@net.bio.net Distribution: world Message-ID: <00995141.83609780.1998@vax.grc.nia.nih.gov> NNTP-Posting-Host: net.bio.net Does anyone know of a paper or report where a G protein coupled receptor muatation was created such that after expression it would produce a dominant negative phenotype. I am particularly interested in peptide receptors but any other information will also be greatly appreciated. Thank you! Chahrzad Montrose Email: chahrzad@vax.grc.nia.nih.gov From owner-7tms_r@net.bio.net Fri Aug 18 23:00:00 1995 Path: biosci!agate!overload.lbl.gov!news.kreonet.re.kr!news.dacom.co.kr!newsfeed.internetmci.com!news.sprintlink.net!in2.uu.net!news3.digex.net!digex.net!not-for-mail From: mmax@universe.digex.net (Max) Newsgroups: bionet.molbio.proteins.7tms_r Subject: GTPase rates of G-proteins Date: 19 Aug 1995 09:17:50 -0400 Organization: Ferret House Lines: 7 Message-ID: <414o9u$r1j@universe.digex.net> NNTP-Posting-Host: universe.digex.net Anyone out there know the relative GTPase rates of the various G-proteins or a reference that might discuss this? Max From owner-7tms_r@net.bio.net Sat Aug 19 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor subtypes Date: 20 Aug 1995 14:11:54 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 61 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508202109.RAA26787@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net Dave Bylund offered this summary of receptor subtype definition: > > Historically, the most useful criteria for defining types and subtypes > of receptors have been pharmacological, i.e. based on the rank order of > potencies of agonists and the relative affinities of antagonists. Other > criteria such as anatomical localization (pre- vs post-synaptic) or > functional response (excitatory vs inhibitory) have not proven to be > useful. More recently, the predicated primary amino acid sequence from > cloning and the relative sequence similarities of related receptors have > also become useful. My hierarchical method for classifying receptors > with examples from the adrenergic receptor family is as follows: > > Superfamily G-protein coupled; Ligand gated ion channels > Family (orClass) Adrenergic > Type (or Subfamily) Alpha-1; Alpha-2; Beta > Subtype(or member) Beta-1;Beta-2; Beta-3; Alpha-2A; Alpha-2B; Alpha-2C > Pharmacological subtypes Alpha-2A and Alpha-2D > > Each level further down, the lower the degree of selectivity of agonists > and antagonist, and the greater the sequence similarity. This raises another question, which is whether 7tms_r that differ in sequence only but have identical ligand specificities can be considered true subtypes. For example, the angiotensin receptor subtypes AT1a and AT1b differ by a few percent in sequence, but there is no convincing difference in ligand speicificity or other properties (rate of desensitization etc.). On the other hand, we published evidence for pharmacological hetereogeneity of AT1 receptors, but our results were ignored by subtype classifiers because we lacked the clone (our group is still trying to clone it). There are many other examples, both of sequence heterogeneity without pharmacological hetereogeneity, and the converse, a distinct pharmacological subtype without a corresponding nucleotide sequence. I suggest that if alternative sequences are found which do not affect the recognition properties of the receptor, then these should be considered something other than a subtype --perhaps an "isoreceptor". If pharmacological differences can be found at a later date, it could be reclassified as a subtype. This proposal is in analogy to the term "isozymes", (which can actually differ substantially from each other without being considered distinct ezymes). -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* --QAA26647.808952075/slc5.INS.CWRU.Edu-- ------------ End of forwarded message -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* From owner-7tms_r@net.bio.net Sat Aug 19 23:00:00 1995 Path: biosci!RECEPTOR.MGH.HARVARD.EDU!lfk From: lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor subtypes Date: 20 Aug 1995 16:14:28 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 108 Sender: daemon@net.bio.net Distribution: world Message-ID: <9508202304.AA10903@receptor> References: <199508202109.RAA26787@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net On 20 Aug 1995 14:11:54 -0700, pre@po.cwru.edu said: > Dave Bylund offered this summary of receptor subtype definition: >> Historically, the most useful criteria for defining types and subtypes >> of receptors have been pharmacological, i.e. based on the rank order of >> potencies of agonists and the relative affinities of antagonists. [points deleted] >> More recently, the predicated primary amino acid sequence from >> cloning and the relative sequence similarities of related receptors >> have also become useful. >> My hierarchical method for classifying receptors >> with examples from the adrenergic receptor family is as follows: >> >> Superfamily G-protein coupled; Ligand gated ion channels >> Family (orClass) Adrenergic >> Type (or Subfamily) Alpha-1; Alpha-2; Beta >> Subtype(or member) Beta-1;Beta-2; Beta-3; Alpha-2A; Alpha-2B; Alpha-2C >> Pharmacological subtypes Alpha-2A and Alpha-2D An evolutionary perspective would be useful here as we are talking about gene duplications and gene diversion. IUPHAR likes a classification based on pharmacology, but there are some surprises if one looks at the evolution of genes. In my analyses I use the following terminology: A ``super-family'' is a collection of protein families that share functional and or structural characteristics, but are not obviously related by sequence similarity. The characteristics for the G-protein--coupled receptor super-family members are: 1) a polypeptide containing seven predicted hydrophobic transmembrane segments (TMS) and 2) the ability to stimulate the exchange of bound GDP for GTP on associated G-protein alpha subunits in response to agonist binding. Super-families are logically subdivided into families. A GCR ``family'' is a collection of receptors whose protein sequences share significant similarity (i.e. >= 20% sequence identity over the predicted TMS). This similarity can be interpreted in an evolutionary context as a group of proteins that share a single common ancestor. A ``group'' refers to receptors that are within a family and also share a common biochemical properties. A pair of additional terms also useful in the classification of molecular data are orthologue and paralogue. A pair of genes are orthologous when they are different because of a speciation event, and are paralogous when the difference resulted from a gene duplication event. > This raises another question, which is whether 7tms_r that differ in > sequence only but have identical ligand specificities can be > considered true subtypes. > For example, the angiotensin receptor subtypes AT1a and > AT1b differ by a few percent in sequence, but there is no convincing > difference in ligand speicificity or other properties (rate of > desensitization etc.). These genes can be explained as recently duplicated genes (thus paralogues) and are on their way via mutations to become distinct genes. I would bet that if knock-outs of these genes are made they individually would have no apparent phenotype. In fact, aren't there a different number of AT1 genes in rat versus mouse versus human (I seem to recall)? > On the other hand, we published evidence for pharmacological > hetereogeneity of AT1 receptors, but our results were ignored by > subtype classifiers because we lacked the clone (our group is still > trying to clone it). > There are many other examples, both of sequence heterogeneity > without pharmacological hetereogeneity, and the converse, a distinct > pharmacological subtype without a corresponding nucleotide sequence. There are plenty of examples of distinct sequence and identical pharmacology (see Reppert's work on the melatonin receptors Mel-1A, Mel-1B, and Mel-1C). But it also depends on the pharmacological probes used. If there are a large number of reagents, I would think that you would increase the probability of finding diverse behavior. I think that there are a myriad of ways to explain the reverse, different beta, gammas, kinases and down stream elements. The recent knock-outs of the En-1 and En-2 genes and the replacement and rescue of En-1 with En-2 are the beginning of a new era in assessment of gene function when there are subtypes. > I suggest that if alternative sequences are found which do not > affect the recognition properties of the receptor, then these should > be considered something other than a subtype --perhaps an > "isoreceptor". If pharmacological differences can be found at a > later date, it could be reclassified as a subtype. This proposal is > in analogy to the term "isozymes", (which can actually differ > substantially from each other without being considered distinct > enzymes). Again, I think that this is ignoring the evolutionary pathway. These genes are present because a gene duplication was found to be useful in a selection. Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Sun Aug 20 23:00:00 1995 Path: biosci!UCHSC.EDU!David.Port From: David.Port@UCHSC.EDU ("Dave Port") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang IV receptors Date: 21 Aug 1995 10:55:29 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508211756.LAA29579@essex.UCHSC.edu> NNTP-Posting-Host: net.bio.net Ang IV receptors Recently, I asked if anyone knew of cells null for Ang II receptors. Paul Ernsberger replied that something to consider was Ang IV receptors and the necessity of using protease inhibitors to prevent formation of Ang IV. Are there references for this out in the literature? I would have sent this to Paul directly, but I couldn't get his e-mail address to work. Thanks David Port port_d@defiance.hsc.colorado.edu From owner-7tms_r@net.bio.net Sun Aug 20 23:00:00 1995 Path: biosci!UNMCVM.UNMC.EDU!DBYLUND From: DBYLUND@UNMCVM.UNMC.EDU ("David B. Bylund --UNMCVM", DBYLUND) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Isoreceptors Date: 21 Aug 1995 10:02:22 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508211702.KAA21678@net.bio.net> NNTP-Posting-Host: net.bio.net FROM: David B. Bylund --UNMCVM(DBYLUND) Paul Ernsberger suggested that the term "isoreceptor" (or perhaps isoceptor) be used to identify "subtypes" that differ in sequence but not in pharmacological characteristics. I would discourage the use of that term since it could just as easily be applied to what I call a subtype (or member) or to what I call pharmacological subytpes. Thus, I use the term "genetic subtypes" for the situation described by Paul. This term has the advantage of parallel construction to the term "pharmacological subtypes" (orthologues with significantly different pharacological characteristics), but also has disadvantages. I would be open to other suggestions--any ideas? David B. Bylund, Ph.D. Department of Pharmacology University of Nebraska Medical Center P.O. Box 986260 Omaha, NE 68198-6260 voice: 402/559-4788 fax: 402/559-7495 EMAIL: DBYLUND@UNMC.EDU From owner-7tms_r@net.bio.net Sun Aug 20 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: skapas@hgmp.mrc.ac.uk (Dr. S Kapas) Newsgroups: bionet.molbio.proteins.7tms_r Subject: . Date: 21 Aug 1995 13:54:07 +0100 Lines: 2 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <419vlf$h17@mserv1.dl.ac.uk> content-length: 2 Original-To: 7tms_r@dl.ac.uk From owner-7tms_r@net.bio.net Sun Aug 20 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!Germany.EU.net!news.dfn.de!fu-berlin.de!cs.tu-berlin.de!fauern!lrz-muenchen.de!ipp-garching.mpg.de!muecke.biochem.mpg.de!krasel From: krasel@muecke.biochem.mpg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GTPase rates of G-proteins Date: 21 Aug 1995 17:41:10 GMT Organization: Rechenzentrum der Max-Planck-Gesellschaft in Garching Lines: 16 Message-ID: <41agfm$1eu0@sat.ipp-garching.mpg.de> References: <414o9u$r1j@universe.digex.net> NNTP-Posting-Host: muecke.biochem.mpg.de X-Newsreader: TIN [version 1.1 PL8] Max (mmax@universe.digex.net) wrote: : Anyone out there know the relative GTPase rates of the various G-proteins : or a reference that might discuss this? I think Gilman has devoted a paragraph to this issue in his 1987 review in Annual Reviews of Biochemistry. However, it distinguishes only between Gi, Go, Gs and Gq (as far as I can remember); nothing about Gz and nothing about cloned isoforms (e.g. Gi-alpha1, -2, ...), which is not surprising since these were not known at this time. --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 Mon Aug 21 23:00:00 1995 Path: biosci!bcm!pendragon.jsc.nasa.gov!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GTPase rates of G-proteins Date: 22 Aug 1995 17:03:58 GMT Organization: University of Michigan Lines: 26 Message-ID: <41d2lu$cr@lastactionhero.rs.itd.umich.edu> References: <414o9u$r1j@universe.digex.net> <41agfm$1eu0@sat.ipp-garching.mpg.de> NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) krasel@muecke.biochem.mpg.de (Cornelius Krasel) wrote: >Max (mmax@universe.digex.net) wrote: >: Anyone out there know the relative GTPase rates of the various G-proteins >: or a reference that might discuss this? > >I think Gilman has devoted a paragraph to this issue in his 1987 review >in Annual Reviews of Biochemistry. However, it distinguishes only between >Gi, Go, Gs and Gq (as far as I can remember); nothing about Gz and nothing >about cloned isoforms (e.g. Gi-alpha1, -2, ...), which is not surprising >since these were not known at this time. > >--Cornelius. One point to keep in mind in thinking about this is that most measurements of GTPase rates are actually measuring GDP release rates. At steady state, the rate-limiting step in the GTPase cycle is the dissociation of GDP. The actual GTPase rate can only be determined by single turnover studies. _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 From owner-7tms_r@net.bio.net Mon Aug 21 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Ang IV receptors Date: 22 Aug 1995 19:49:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 70 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508230246.WAA24782@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net I am posting this in case anyone else is interested: > Ang IV receptors > Recently, I asked if anyone knew of cells null for Ang II receptors. Paul > Ernsberger replied that something to consider was Ang IV receptors and the > necessity of using protease inhibitors to prevent formation of Ang IV. Are > there references for this out in the literature? > 1). Jarvis MF, Gessner GW, Ly CQ: The angiotensin hexapeptide 3-8 fragment potently inhibits [125I]angiotensin II binding to non-AT1 or -AT2 recognition sites in bovine adrenal cortex. Eur J Pharmacol 219:319-322, 1992 2). Swanson GN, Hanesworth JM, Sardinia MF, Coleman JKM, Wright JW, Hall KL, Miller-Wing AV, Stobb JW, Cook VI, Harding EC, Harding JW: Discovery of a distinct binding site for angiotensin II (3-8), a putative angiotensin IV receptor. Regul Pept 40:409-419, 1992 3). Hall KL, Hanesworth JM, Ball AE, Felgenhauer GP, Hosick HL, Harding JW: Identification and characterization of a novel angiotensin binding site in cultured vascular smooth muscle cells that is specific for the hexapeptide (3-8) fragment of angiotensin II, angiotensin IV. Regul Pept 44:225-232, 1993 4). Hanesworth JM, Sardinia MF, Krebs LT, Hall KL, Harding JW: Elucidation of a specific binding site for angiotensin II(3-8), angiotensin IV, in mammalian heart membranes. J Pharmacol Exp Ther 266:1036-1042, 1993 5). Miller-Wing AV, Hanesworth JM, Sardinia MF, Hall KL, Wright JW, Speth RC, Grove KL, Harding JW: Central angiotensin IV binding sites: Distribution and specificity in guinea pig brain. J Pharmacol Exp Ther 266:1718-1726, 1993 6). Sardinia MF, Hanesworth JM, Krebs LT, Harding JW: AT4 receptor binding characteristics: D-amino acid- and glycine-substituted peptides. Peptides 14:949-954, 1993 7). Bernier SG, Fournier A, Guillemette G: A specific binding site recognizing a fragment of angiotensin II in bovine adrenal cortex membranes. Eur J Pharmacol 271:55-63, 1994 8). Dulin NO, Ernsberger P, Suciu DJ, Douglas JG: Rabbit renal epithelial angiotensin II receptors. Am J Physiol Renal,Fluid Electrolyte Physiol 267:F776-F782, 1994 9). Harding JW, Wright JW, Swanson GN, Hanesworth JM, Krebs LT: AT4 receptors: Specificity and distribution. Kidney Int 46:1510-1512, 1994 10). N„veri L, Str”mberg C, Saavedra JM: Angiotensin IV reverses the acute cerebral blood flow reduction after experimental subarachnoid hemorrhage in the rat. J Cereb Blood Flow Metab 14:1096-1099, 1994 11). Sardinia MF, Hanesworth JM, Krishnan F, Harding JW: AT4 receptor structure-binding relationship: N-terminal-modified angiotensin IV analogues. Peptides 15:1399-1406, 1994 12). Wright JW, Harding JW: Brain angiotensin receptor subtypes in the control of physiological and behavioral responses. Neurosci Biobehav Rev 18:21-53, 1994 13). Wang L, Eberhard M, K”hler E, Erne P: A specific binding site for angiotensin II(3-8), angiotensin IV, in rabbit cardiac fibroblasts. J Recept Res 15:517-527, 1995 14). Wang L, Eberhard M, Erne P: Stimulation of DNA and RNA synthesis in cultured rabbit cardiac fibroblasts by angiotensin IV. Clin Sci 88:557-562, 1995 -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* From owner-7tms_r@net.bio.net Mon Aug 21 23:00:00 1995 Path: biosci!bcm!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!tank.news.pipex.net!pipex!swrinde!emory!usenet From: medtjm@bimcore.emory.edu (T. J. Murphy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Isoreceptors (& evotypes) Date: 22 Aug 1995 20:37:17 GMT Organization: Biomolecular Computing Resource, Emory University Lines: 26 Distribution: world Message-ID: <41df5t$a18@emory.mathcs.emory.edu> References: <199508211702.KAA21678@net.bio.net> Reply-To: medtjm@bimcore.emory.edu NNTP-Posting-Host: bimcore.emory.edu I simply call the rat and mouse AT1a and AT1b receptors *isoforms*. In lighter moments, I call them snapshots of evolution in progress (evotypes???). ;) They are a pretty good example of an isolated instance of gene duplication, likely unique to the branch upon which little laboratory rodents bloom. Could've happened to any gene in any species at any time, but it just so happened here. Given that humans and other species have only one AT1 receptor gene, it's hard to argue for AT1 receptor subtypes. What would you call the human gene then? It would have to be AT1a/b because by sequence it plops down right in the middle of the AT1a and AT1b. That, of course, would be ludicrous. The simplest solution is to utilize asterisks copiously and footnote these kinds of oddities as isoforms. In the meantime, we can only wait for a similar genetic hiccup somewhere in our fair species. Would that he/she have many children, that they in turn are fruitful and multiply, etc, etc, etc. --- TJ Murphy Asst. Professor Dept of Pharmacology Emory University School of Medicine From owner-7tms_r@net.bio.net Mon Aug 21 23:00:00 1995 Path: biosci!bcm!pendragon.jsc.nasa.gov!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor subtypes Date: 22 Aug 1995 14:26:17 GMT Organization: University of Michigan Lines: 30 Message-ID: <41cpe9$p16@lastactionhero.rs.itd.umich.edu> References: <199508202109.RAA26787@slc5.INS.CWRU.Edu> <9508202304.AA10903@receptor> NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) wrote: > >> For example, the angiotensin receptor subtypes AT1a and >> AT1b differ by a few percent in sequence, but there is no convincing >> difference in ligand speicificity or other properties (rate of >> desensitization etc.). > >These genes can be explained as recently duplicated genes (thus >paralogues) and are on their way via mutations to become distinct >genes. I would bet that if knock-outs of these genes are made they >individually would have no apparent phenotype. In fact, aren't there a >different number of AT1 genes in rat versus mouse versus human (I seem >to recall)? > The one situation where this would not hold true is if the AT1a and AT1b are expressed in different tissues. Then a knockout wouldn't change the pharmacology but would change the tissue distribution of responses. That alone could produce a striking phenotype. See JBC 270:18719 which shows that the AT1a knockout does indeed have a striking phenotype and suggests that the AT1b may have a less dominant role. _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 From owner-7tms_r@net.bio.net Tue Aug 22 23:00:00 1995 Path: biosci!CELLBIO.WUSTL.EDU!kblumer From: kblumer@CELLBIO.WUSTL.EDU (Ken Blumer) Newsgroups: bionet.molbio.proteins.7tms_r Subject: 7TMR's w/out proline in TM6 Date: 23 Aug 1995 15:51:55 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net 7TMers-Many 7TMR's have a proline in TM6. Are there 7TMR's that are exceptions to this rule? --Ken From owner-7tms_r@net.bio.net Tue Aug 22 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!usenet.eel.ufl.edu!newsfeed.internetmci.com!tank.news.pipex.net!pipex!in1.uu.net!world!news From: rrohan@world.std.com (Richard Rohan) Subject: Re: GTPase rates of G-proteins Message-ID: Sender: news@world.std.com (Mr Usenet Himself) Nntp-Posting-Host: world.std.com Organization: The World @ Software Tool & Die X-Newsreader: Forte Agent .99.82 References: <414o9u$r1j@universe.digex.net> <41agfm$1eu0@sat.ipp-garching.mpg.de> Date: Thu, 24 Aug 1995 04:59:37 GMT Lines: 17 Max (mmax@universe.digex.net) wrote: >: Anyone out there know the relative GTPase rates of the various G-proteins >: or a reference that might discuss this? Two references mentioned in a recent review by EJ Neer (Cell 80:249-57 1995) are DJ Carty et al (JBC 265:6268-73 1990) and ME Linder et al. (JBC 265:8243-51 1990). This topic was also covered by HR Bourne & L Stryer (Nature 358:541-3 1992). HTH --- Rich Rohan, PhD (rrohan@world.std.com) Independent Molecular Biology Consultant to the Internet Moderator REPRENDO@world.std.com - Reproductive Endocrinology List From owner-7tms_r@net.bio.net Wed Aug 23 23:00:00 1995 Path: biosci!HELIX.NIH.GOV!tibonner From: tibonner@HELIX.NIH.GOV (Tom Bonner) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMR's w/out proline in TM6 Date: 24 Aug 1995 06:32:21 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net >7TMers-Many 7TMR's have a proline in TM6. Are there 7TMR's that are >exceptions to this rule? >--Ken The orphan receptor RTA and the second parathyroid hormone receptor (PTH2). Tom I. Bonner, Lab of Cell Biology, National Institute of Mental Health Bldg. 36 Room 3A-07, Bethesda MD 20892-4090 Telephone: 301-496-8907, FAX 301-402-1748 tibonner@helix.nih.gov From owner-7tms_r@net.bio.net Wed Aug 23 23:00:00 1995 Path: biosci!MIRIS.MED.YALE.EDU!mike From: mike@MIRIS.MED.YALE.EDU ("Michael Singer") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 7TMR's w/out proline in TM6 Date: 24 Aug 1995 06:36:49 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <9508240938.ZM18683@miris.med.yale.edu> NNTP-Posting-Host: net.bio.net Hi Ken, Most olfactory GPCRs do not have prolines in TM6. Michael Singer Section of Neurobiology Yale University School of Medicine -------------------------------------------------------------------------------- 7TMers-Many 7TMR's have a proline in TM6. Are there 7TMR's that are exceptions to this rule? --Ken From owner-7tms_r@net.bio.net Sun Aug 27 23:00:00 1995 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.reston.ans.net!newsfeed.internetmci.com!bloom-beacon.mit.edu!news5.ner.bbnplanet.net!news3.near.net!yale!yale!oitnews.harvard.edu!bloch.nmr.mgh.harvard.edu!nntp.mgh.harvard.edu!lfk From: lfk@receptor.mgh.harvard.edu (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Excerpted TOC from 9/15 JBC Date: 28 Aug 1995 17:33:04 GMT Organization: Mass. General Hospital, Renal Unit Lines: 40 Distribution: world Message-ID: NNTP-Posting-Host: receptor.mgh.harvard.edu AU Wayman-G-A. Impey-S. Storm-D-R. TI Ca2+ inhibition of type III adenylyl cyclase in vivo SO J-Biol-Chem. 1995 September 15. 270(37). p 21480. AU Nanevicz-T. Ishii-M. Wang-L. Chen-M. Chen-J. Turck-C-W. Cohen-F-E. Coughlin-S-R. TI Mechanisms of thrombin receptor agonist specificity. Chimeric receptors and complementary mutations identify an agonist recognition site SO J-Biol-Chem. 1995 September 15. 270(37). p 21619. AU Creasy-C-L. Chernoff-J. TI Cloning and characterization of a human protein kinase with homology to Ste20 SO J-Biol-Chem. 1995 September 15. 270(37). p 21695. AU Zhang-R. Cai-H. Fatima-N. Buczko-E. Dufau-M-L. TI Functional glycosylation sites of the rat luteinizing hormone receptor required for ligand binding SO J-Biol-Chem. 1995 September 15. 270(37). p 21722. AU Ray-K. Kunsch-C. Bonner-L-M. Robishaw-J-D. TI Isolation of cDNA clones encoding eight different human G protein {gamma} subunits, including three novel forms designated the {gamma}4, {gamma}10, and {gamma}11 subunits SO J-Biol-Chem. 1995 September 15. 270(37). p 21765. AU Gorczyca-W-A. Polans-A-S. Surgucheva-I-G. Subbaraya-I. Baehr-W. Palczewski-K. TI Guanylyl cyclase activating protein. A calcium-sensitive regulator of phototransduction SO J-Biol-Chem. 1995 September 15. 270(37). p 22029. -- Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Tue Aug 29 23:00:00 1995 Path: biosci!MFOUR.MED.KYOTO-U.AC.JP!hiratamz From: hiratamz@MFOUR.MED.KYOTO-U.AC.JP (Hirata, Masakazu) Newsgroups: bionet.molbio.proteins.7tms_r Subject: MnCl2 and G-protein Date: 30 Aug 1995 21:14:51 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <199508310401.NAA06069@mfour.mfour.med.kyoto-u.ac.jp> NNTP-Posting-Host: net.bio.net Hello, Does anybody know how MnCl2 affect G-proteins? Is GTP hydrolysis reduced by manganese ion? Is the effect differnt from Mg2+? Thank you very much for your consideration. Masakazu Hirata, MD Kyoto University Faculty of Medicine From owner-7tms_r@net.bio.net Thu Aug 31 23:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!nctuccca.edu.tw!aladdin.iii.org.tw!NewsWatcher!user From: hungkaic@dtk1.ym.edu.tw (Hung-Kai Chen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GTPase rates of G-proteins Date: Sat, 02 Sep 1995 02:01:00 +0800 Organization: National Yang-Ming University Lines: 30 Message-ID: References: <414o9u$r1j@universe.digex.net> <41agfm$1eu0@sat.ipp-garching.mpg.de> NNTP-Posting-Host: 140.129.70.95 In article <41agfm$1eu0@sat.ipp-garching.mpg.de>, krasel@muecke.biochem.mpg.de (Cornelius Krasel) wrote: >Max (mmax@universe.digex.net) wrote: >: Anyone out there know the relative GTPase rates of the various G-proteins >: or a reference that might discuss this? > >I think Gilman has devoted a paragraph to this issue in his 1987 review >in Annual Reviews of Biochemistry. However, it distinguishes only between >Gi, Go, Gs and Gq (as far as I can remember); nothing about Gz and nothing >about cloned isoforms (e.g. Gi-alpha1, -2, ...), which is not surprising >since these were not known at this time. > >--Cornelius. > Cornelius Krasel, email: krasel@alf.biochem.mpg.de In general, the rate of GTPase in an in vitro assay is relatively slow, ranging from12 to 250 mmol/min/mol of protein (Proc. Natl. Acad. Sci.1984, 81:5704-5708). For some other GTPase: Mx1 protein: 6/mol/min/mol of protein (J Biol. Chem. 1991. 266(32):21409-21415.) Dynamin: 1/mol/min/mol of protein (J. Cell Sci. 1991. 14:143-145.) GTPase in vesicular transport (Annu. Rev. Biochem. 1994. 63:949-90) Hung-Kai Chen --------------------------------------------------------------------------- Hung-Kai Chen Email: hungkaic@dtk1.ym.edu.tw Institute of Microbiology & Immunology, National Yang-Ming University, Taipei, Taiwan, R.O.C.