From owner-7tms_r@net.bio.net Fri Mar 03 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!agate!howland.reston.ans.net!paladin.american.edu!newsfeed.ACO.net!swidir.switch.ch!univ-lyon1.fr!ghost.dsi.unimi.it!genes!karami From: karami@genes.icgeb.trieste.it (Ali Karami) Subject: Mab for OspA of Borrelia???? Message-ID: <1995Mar3.181631.6123@genes.icgeb.trieste.it> Organization: ICGEB X-Newsreader: TIN [version 1.2 PL2] Date: Fri, 3 Mar 1995 18:16:31 GMT Lines: 27 From karami Fri Mar 3 18:58:51 1995 Return-Path: Received: by icgeb.trieste.it (AA05218); Fri, 3 Mar 95 18:58:50 +0100 Date: Fri, 3 Mar 95 18:58:50 +0100 From: Ali Karami Message-Id: <9503031758.AA05218@icgeb.trieste.it> To: karami@icgeb.trieste.it (Ali Karami) Subject: Re: Mab for OspA of Borrelia???? Newsgroups: bionet.immunology Organization: ICGEB X-Newsreader: TIN [version 1.2 PL2] Status: OR In article <1995Mar3.174607.5025@genes.icgeb.trieste.it> you wrote: I need Monoclonal Ab against OspA of Borrelia burgdorferi ? How can I find it ? I want to do few Immuno blotting with mY protein. I would appreciate Any help. Thank you Ali Karami Ali@biobase.dk From owner-7tms_r@net.bio.net Sat Mar 04 22:00:00 1995 Path: biosci!GIBBS.OIT.UNC.EDU!laiter From: laiter@GIBBS.OIT.UNC.EDU (Sergei Laiter) Newsgroups: bionet.molbio.proteins.7tms_r Subject: models of B2 adrenoreceptors Date: 5 Mar 1995 11:58:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear netters, I'm modeling G-protein coupled receptors and I'mparticularly interested in existing models of B2 adrenoreceptors. If anyone would provide me with their models of these receptors I would appreciate it. -Sergei From owner-7tms_r@net.bio.net Sun Mar 05 22:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.nysernet.net!news.sprintlink.net!pipex!warwick!news.dcs.warwick.ac.uk!hgmp.mrc.ac.uk!daresbury!not-for-mail From: mcroos@imb-jena.de (Martin Roos) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Mab for OspA of Borrelia???? Date: 6 Mar 1995 09:37:00 -0000 Lines: 19 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3jel3s$lik@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Ali Karami wrote: >I need Monoclonal Ab against OspA of Borrelia burgdorferi ? > > How can I find it ? I want to do few Immuno blotting with mY protein. I >would appreciate Any help. You might succeed in getting a Mab by contacting either Michael Kramer (Universitdt Heidelberg, Institut f|r Immunologie, Abteilung f|r Immunpathologie) or Markus Simon (Freiburg, Max-Planck.Institut f|r Immunbiologie). In the lab of Michael Kramer I once worked as a trainee. If you will not find out the phone numbers just contact me again. Greetings Martin Roos mcroos@imb-jena.de From owner-7tms_r@net.bio.net Tue Mar 07 22:00:00 1995 Path: biosci!NIFTYSERVE.OR.JP!JCF11024 From: JCF11024@NIFTYSERVE.OR.JP (=?ISO-2022-JP?B?GyRCNmJFRCEhPSEwbE86GyhC?=) Newsgroups: bionet.molbio.proteins.7tms_r Subject: HELP Date: 8 Mar 1995 06:32:12 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <199503081432.XAA10497@inetnif.niftyserve.or.jp> NNTP-Posting-Host: net.bio.net HELP From owner-7tms_r@net.bio.net Tue Mar 07 22:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!gatech!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r,bionet.molbio.proteins Subject: Ultracold storage Date: 9 Mar 1995 00:01:21 GMT Organization: University of Michigan Lines: 15 Message-ID: <3jlggi$41c@lastactionhero.rs.itd.umich.edu> NNTP-Posting-Host: pm002-24.dialip.mich.net Xref: biosci bionet.molbio.proteins.7tms_r:199 bionet.molbio.proteins:3929 We just got a new freezer that can go to -86 C and I wondered if anybody had personal experience with storage of G proteins and/or 7tm receptors/membranes at different temperatures. We've always stored them at -70. Do they remain more stable at lower temps (e.g. -80 or -86). My initial reaction was "put it as cold as it gets" but apparently that leads to more problems with frosting up and more stress on motors and compressors. So - at what temperature do you all store your precious G proteins and 7tms_r's and is there any rational basis for the choice? Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Wed Mar 08 22:00:00 1995 Path: biosci!SMTPGATE.ICOS.COM!pgray From: pgray@SMTPGATE.ICOS.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 9 Mar 1995 13:03:59 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <9502097947.AA794783079@smtpgate.icos.com> NNTP-Posting-Host: net.bio.net Subscribe From owner-7tms_r@net.bio.net Tue Mar 14 22:00:00 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: subscribe Date: 15 Mar 1995 09:25:19 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <9503151345.AA02401@receptor> References: <9502097947.AA794783079@smtpgate.icos.com> NNTP-Posting-Host: net.bio.net Dear Dr. Gray, We do not have any informative data about your clone R2 yet. We did start a newsgroup and a mailing list under the BIOSCI system. If you are able to read any of the electronic newsgroups bionet.*, then just ask the administrator of that system to add the bionet.molbio.proteins.7tms_r newsgroup. If you are unable to read news and want to get it by email send a message to biosci-server@net.bio.net in the message say 'subscribe 7tms_r' Then all email to 7tms_r@net.bio.net willl come to your email address and also any message you send to 7tms_r@net.bio.net will also come to your address. Best wishes, also look at GCRDbWWW on the World Wide Web http://receptor.mgh.harvard.edu/GCRDBHOME.html Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-735-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Thu Mar 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: Associate Scientist Position Opening Date: 17 Mar 1995 09:47:00 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 43 Sender: daemon@net.bio.net Distribution: world Message-ID: <9503172046.AA5699@pho018.sb.com> NNTP-Posting-Host: net.bio.net Dear 7TM Netters, I hope this is an appropriate use of the net. My apologies if I am out of line. I am seeking a motivated scientist to join an ongoing multidisciplinary effort to investigate the structure and ligand interactions of bioactive peptide receptors. Requirements include a BS and MS in biochemistry, pharmacology or a related field. Experience in receptor characterization (ligand binding and signal transduction assays) and familiarity with data analysis programs is preferred. Additional experience in molecular biology (subcloning, mutagenesis, PCR, sequencing, and gene expression) and the operation of laboratory robots is desirable. Examples of recent work from the laboratory can be found in the following references. Lee, J.A., Elliott, Sutiphong, J.A, Friesen, W.J., Ohlstein, E.H. Stadel, J.M., Gleason, J.G., and Peishoff, C. E. (1994) "Tyrosine 129 Is Important to the Peptide Affinity and Selectivity of Human Endothelin A Receptor" Proc. Natl. Acad. Sci. USA 91, 7164-7168. Lee, J.A., Sutiphong, J.A., Longton, E.D., Peishoff, C. E., Stadel, J.M., Kumar, C., Ohlstein, E.H., Gleason, J.D., and Elliott, J.D (1994) "Lysine 182 of the Endothelin B Receptor is Important for Agonist Selectivity and Antagonist Affinity: Evidence for the Overlap of Peptide and Non-peptide Ligand Binding Sites" Biochemistry 33, 14543-14549. For consideration, please contact Dr. Jonathan Lee. Jonathan A. Lee SmithKline Beecham Pharmaceuticals Dept. Macromolecular Science Mail COde UE 0447 709 Swedeland Road King of Prussia, PA 19406 TEL: 610-270-7588 FAX: 610-270-4091 Email: Jonathan A Lee%notes@ab.com From owner-7tms_r@net.bio.net Sun Mar 19 22:00:00 1995 Path: biosci!daresbury!not-for-mail From: mcroos@imb-jena.de (Martin Roos) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Blotting of membrane receptors Date: 20 Mar 1995 15:25:12 -0000 Lines: 31 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3kk6oo$8fn@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Dear netters, my actual lab problem: How can I increase the semi-dry transfer of a membran receptor (here: endothelin-receptor) from SDS-PAGE to PVDF membranes? I am currently using a continous CAPS-buffered system, and I played around with the SDS and MeOH content on the cathodic site of the blot sandwich, but the results were not very convincing. Any suggestions for the right proportion of SDS/MeOH? Any suggestion for another blot system (acidic) ? I will be happy about any comment. Ciao, Martin ______________________________________________________________________________ Martin Roos.................................. e-mail: mcroos@imb-jena.de Institut f|r Molekulare Biotechnologie e.V... fax: ++49-3641-656431 Proteinlabor ................................ tel: ++49-3641-656263 Beutenbergstr. 11.......................................................... D-07745 Jena............................................................... Germany _______________________________________________________________________________ From owner-7tms_r@net.bio.net Sun Mar 19 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: Associate Scientist Position Opening Date: 20 Mar 1995 07:55:10 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: <9503201855.AA6337@pho018.sb.com> NNTP-Posting-Host: net.bio.net Dear 7TM Netters, I had a typographical error on my message concerning the Associate Scientist position at SmithKline Beecham. My E MAil address is : Jonathan A. Lee SmithKline Beecham Pharmaceuticals Dept. Macromolecular Science Mail COde UE 0447 709 Swedeland Road King of Prussia, PA 19406 TEL: 610-270-7588 FAX: 610-270-4091 Email: Jonathan A Lee%notes@sb.com (NOT ab.com) From owner-7tms_r@net.bio.net Mon Mar 20 22:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!news.sprintlink.net!pipex!sunic!sunic.sunet.se!news.funet.fi!news.csc.fi!news.helsinki.fi!kruuna!paakkone From: paakkone@cc.Helsinki.FI (Kimmo Paakkonen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 17 Mar 1995 13:11:53 GMT Organization: University of Helsinki Lines: 1 Message-ID: <3kc1qp$a88@oravannahka.Helsinki.FI> NNTP-Posting-Host: kruuna.helsinki.fi Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Newsreader: TIN [version 1.2 PL0] subscribe From owner-7tms_r@net.bio.net Thu Mar 23 22:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!cs.utexas.edu!convex!news.duke.edu!news-feed-1.peachnet.edu!insosf1.infonet.net!newshost.marcam.com!zip.eecs.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Mol Pharm Gordon Conference Date: 24 Mar 1995 21:12:23 GMT Organization: University of Michigan Lines: 13 Message-ID: <3kvcjn$gb8@lastactionhero.rs.itd.umich.edu> NNTP-Posting-Host: warbler.med.umich.edu Hi 7tms_r's, Things have been pretty quiet here so I'll try to start something. Did anyone go to the Molecular Pharmacology Gordon Conference in California in late February? I wanted to go but couldn't. Any exciting new developments? Any word on x'tal structure of G protein beta subunits? Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Sun Mar 26 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: Record-breaking loops? Date: 27 Mar 1995 13:53:35 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 6 Sender: daemon@net.bio.net Distribution: world Message-ID: <9503271656.ZM19321@miris.med.yale.edu> NNTP-Posting-Host: net.bio.net Can anyone name some GPCRs with especially large 2nd extracellular loops (the loops between TM4 and TM5)? How many residues? Thanks, Michael Singer Yale School of Medicine From owner-7tms_r@net.bio.net Mon Mar 27 23:00:00 1995 Path: biosci!WH.BAYER.COM!hyw From: hyw@WH.BAYER.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Re; endocytosis poison Date: 28 Mar 1995 07:29:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 99 Sender: daemon@net.bio.net Distribution: world Message-ID: <9503281522.AA13288@mrcs1> NNTP-Posting-Host: net.bio.net q q q q From owner-7tms_r@net.bio.net Tue Mar 28 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: incomplete removal of radio-labelled agonist by unlabelled agonist? Date: 29 Mar 1995 08:31:51 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 35 Sender: daemon@net.bio.net Distribution: world Message-ID: <26E101BA_000DF7E0.0098E1832746FFF3$27_1@UK.AC.LEEDS.BIO.VAX> NNTP-Posting-Host: net.bio.net Greetings, I wonder if anyone can help to answer a question. The human NK2 receptor: This receptor has approx. nM affinity for its natural peptide and for the non-peptide antagonist SR48,968. If the radio-ligand is 125I-NKA, then the IC50 for unlabelled NKA and SR48,968 is approx nM. In a decoupled system, the IC50 for the NKA drops to the uM range. If the radiolabelled ligand is 3H-SR48,968, then the IC50 for unlabelled SR48,968 is nM, but that for NKA is uM. The latter two IC50 values are unaltered in the decoupled system. Okay, so this can be explained by the ternary model - that is that 125I-NKA highlights a population of "active conformation" coupled-receptors which have a high affinity for both NKA and SR48,968. If a decoupled system is imposed, the affinity of NKA drops but that of SR48,968 is unaltered. However, if 3H-SR48,968 is used as the radio-ligand then a population of predominantly "inactive conformation" decoupled-receptors is highlighted. NKA binds poorly to this state and decoupling conditions have little effect. What I don't understand is the following. When 3H-SR48,968 is used as the radio-ligand, unlabelled NKA can only remove about 70% of the counts compared to unlabelled SR48,968. Why? See "Rosenkilde et al. (1994) J. Biol. Chem. 269, 28160-28164" for the data. Thanks in advance, Dan Donnelly Dept Biochem + Mol Biol, University of Leeds, Leeds LS2 9JT, UK. bmb6hdd@biovax.leeds.ac.uk From owner-7tms_r@net.bio.net Tue Mar 28 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: incomplete removal of radio-labelled ANTagonist by unlabelled agonist? Date: 29 Mar 1995 08:32:27 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 35 Sender: daemon@net.bio.net Distribution: world Message-ID: <26E101BA_000DF390.0098E1833A014FB3$27_2@UK.AC.LEEDS.BIO.VAX> NNTP-Posting-Host: net.bio.net Greetings, I wonder if anyone can help to answer a question. The human NK2 receptor: This receptor has approx. nM affinity for its natural peptide and for the non-peptide antagonist SR48,968. If the radio-ligand is 125I-NKA, then the IC50 for unlabelled NKA and SR48,968 is approx nM. In a decoupled system, the IC50 for the NKA drops to the uM range. If the radiolabelled ligand is 3H-SR48,968, then the IC50 for unlabelled SR48,968 is nM, but that for NKA is uM. The latter two IC50 values are unaltered in the decoupled system. Okay, so this can be explained by the ternary model - that is that 125I-NKA highlights a population of "active conformation" coupled-receptors which have a high affinity for both NKA and SR48,968. If a decoupled system is imposed, the affinity of NKA drops but that of SR48,968 is unaltered. However, if 3H-SR48,968 is used as the radio-ligand then a population of predominantly "inactive conformation" decoupled-receptors is highlighted. NKA binds poorly to this state and decoupling conditions have little effect. What I don't understand is the following. When 3H-SR48,968 is used as the radio-ligand, unlabelled NKA can only remove about 70% of the counts compared to unlabelled SR48,968. Why? See "Rosenkilde et al. (1994) J. Biol. Chem. 269, 28160-28164" for the data. Thanks in advance, Dan Donnelly Dept Biochem + Mol Biol, University of Leeds, Leeds LS2 9JT, UK. bmb6hdd@biovax.leeds.ac.uk From owner-7tms_r@net.bio.net Tue Mar 28 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!Norway.EU.net!EU.net!howland.reston.ans.net!news2.near.net!das-news2.harvard.edu!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: JBC TOC Realted to 7tms_r (3/24-4/7) Date: 29 Mar 1995 12:57:57 GMT Organization: Mass. General Hospital, Renal Unit Lines: 147 Distribution: world Message-ID: NNTP-Posting-Host: receptor.mgh.harvard.edu AU Raychowdhury-M-K. Yukawa-M. Collins-L-J. McGrail-S-H. Kent-K-C. Ware-J-A. TI Alternative splicing produces a divergent cytoplasmic tail in the human endothelial thromboxane A2 receptor. Vol. 269 (1994) 19256-19261. SO J-Biol-Chem. 1995 Mar 24. 270(12). p 7011. AU Yakovlev-A-G. Krueger-K-E. Faden-A-I. TI Structure and expression of a rat {kappa} opioid receptor gene SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6421. AU Justice-J-M. Bliziotes-M-M. Stevens-L-A. Moss-J. Vaughan-M. TI Involvement of N-myristoylation in monoclonal antibody recognition sites on chimeric G protein {alpha} subunits SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6436. AU Kojro-E. Fahrenholz-F. TI Ligand-induced cleavage of the V2 vasopressin receptor by a plasma membrane metalloproteinase SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6476. AU Parker-E-M. Swigart-P. Nunnally-M-H. Perkins-J-P. Ross-E-M. TI Carboxyl-terminal domains in the avian {beta}1-adrenergic receptor that regulate agonist-promoted endocytosis SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6482. AU Wang-J. Ross-E-M. TI The carboxyl-terminal anchorage domain of the turkey {beta}1-adrenergic receptor is encoded by an alternatively spliced exon SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6488. AU Gardella-T-J. Luck-M-D. Wilson-A-K. Keutmann-H-T. Nussbaum-S-R. Potts-J-T-Jr.. Kronenberg-H-M. TI Parathyroid hormone (PTH)-PTH-related peptide hybrid peptides reveal functional interactions between the 1-14 and 15-34 domains of the ligand SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6584. AU Zhang-J. Zhang-J. Benovic-J-L. Sugai-M. Wetzker-R. Gout-I. Rittenhouse-S-E. TI Sequestration of a G-protein {beta}{gamma} subunit or ADP-ribosylation of Rho can inhibit thrombin-induced activation of platelet phosphoinositide 3-kinases SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6589. AU Fukuda-K. Kato-S. Mori-K. TI Location of regions of the opioid receptor involved in selective agonist binding SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6702. AU Greene-N-M. Williams-D-S. Newton-A-C. TI Kinetics and localization of the phosphorylation of rhodopsin by protein kinase C SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6710. AU Morrison-D-F. Ting-T-D. Vallury-V. Ho-Y-K. Crouch-R-K. Corson-D-W. Mangini-N-J. Pepperberg-D-R. TI Reduced light-dependent phosphorylation of an analog visual pigment containing 9-demethylretinal as its chromophore SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6718. AU Ryba-N-J-P. Tirindelli-R. TI A novel GTP-binding protein {gamma}-subunit, G{gamma}8, is expressed during neurogenesis in the olfactory and vomeronasal neuroepithelia SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6757. AU Seckl-M-J. Morii-N. Narumiya-S. Rozengurt-E. TI Guanosine 5{prime}-3-O-(thio)triphosphate stimulates tyrosine phosphorylation of p125FAK and paxillin in permeabilized Swiss 3T3 cells. Role of p21rho. SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6984. AU Iwasaki-S. Homma-T. Matsuda-Y. Kon-V. TI Endothelin receptor subtype B mediates autoinduction of endothelin-1 in rat mesangial cells SO J-Biol-Chem. 1995 Mar 24. 270(12). p 6997. AU Clark-K-J. Murray-A-W. TI Evidence that the bradykinin-induced activation of phospholipase D and of the mitogen-activated protein kinase cascade involve different protein kinase C isoforms SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7097. AU Rousell-J. Haddad-E-B. Mak-J-C-W. Barnes-P-J. TI Transcriptional down-regulation of m2 muscarinic receptor gene expression in human embryonic lung (HEL 299) cells by protein kinase C SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7213. AU Guiramand-J. Montmayeur-J-P. Ceraline-J. Bhatia-M. Borrelli-E. TI Alternative splicing of the dopamine D2 receptor directs specificity of coupling to G-proteins SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7354. AU Hogger-P. Shockley-M-S. Lameh-J. Sadee-W. TI Activating and inactivating mutations in N- and C-terminal i3 loop junctions of muscarinic acetylcholine Hm1 receptors SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7405. AU Pian-M-S. Dobbs-L-G. TI Evidence for G{beta}{gamma}-mediated cross-talk in primary cultures of lung alveolar cells. Pertussis toxin-sensitive production of cAMP. SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7427. AU Chaikof-E-L. Caban-R. Yan-C-N. Rao-G-N. Runge-M-S. TI Growth-related responses in arterial smooth muscle cells are arrested by thrombin receptor antisense sequences SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7431. AU Buggy-J-J. Livingston-J-N. Rabin-D-U. Yoo-Warren-H. TI Glucagonglucagon-like peptide I receptor chimeras reveal domains that determine specificity of glucagon binding SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7474. AU Levin-L-R. Reed-R-R. TI Identification of functional domains of adenylyl cyclase using in vivo chimeras SO J-Biol-Chem. 1995 Mar 31. 270(13). p 7573. AU Pan-Z. Kravchenko-V-V. Ye-R-D. TI Platelet-activating factor stimulates transcription of the heparin-binding epidermal growth factor-like growth factor in monocytes. Correlation with an increased {kappa}B binding activity. SO J-Biol-Chem. 1995 April 7. 270(14). p 7787. AU Jeohn-G-H. Takahashi-K. TI Purification and characterization of a vasoactive intestinal polypeptide-degrading endoprotease from porcine antral mucosal membranes SO J-Biol-Chem. 1995 April 7. 270(14). p 7809. AU Kroog-G-S. Sainz-E. Worland-P-J. Akeson-M-A. Benya-R-V. Jensen-R-T. Battey-J-F. TI The gastrin-releasing peptide receptor is rapidly phosphorylated by a kinase other than protein kinase C after exposure to agonist SO J-Biol-Chem. 1995 April 7. 270(14). p 8217. AU Vouret-Craviari-V. Grall-D. Chambard-J-C. Rasmussen-U-B. Pouyssegur-J. Obberghen-Schilling-E-V. TI Post-translational and activation-dependent modifications of the G protein-coupled thrombin receptor SO J-Biol-Chem. 1995 April 7. 270(14). p 8367. -- Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Tue Mar 28 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!Norway.EU.net!EU.net!Germany.EU.net!zib-berlin.de!cs.tu-berlin.de!fauern!winx03!wrzx15!phak004 From: phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Mol Pharm Gordon Conference Date: 28 Mar 1995 19:08:24 GMT Organization: University of Wuerzburg, Germany Lines: 16 Message-ID: <3l9mr8$ee6@winx03.informatik.uni-wuerzburg.de> References: <3kvcjn$gb8@lastactionhero.rs.itd.umich.edu> NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] Rick Neubig (RNeubig@umich.edu) wrote: > Any exciting new developments? Any word on x'tal structure of > G protein beta subunits? Is there something like this to be expected? (If not, I always wondered how complicated it would be to determine the structure of a single WD40 repeat by NMR?) --Cornelius. PS: Note changed location and email address :-) -- /* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ /* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Tue Mar 28 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: "locked R* receptors" / constitutively active receptors??????? Date: 29 Mar 1995 08:34:04 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 23 Sender: daemon@net.bio.net Distribution: world Message-ID: <26E101BA_000DF7E0.0098E183748AF7D3$28_1@UK.AC.LEEDS.BIO.VAX> NNTP-Posting-Host: net.bio.net Another question: Constitutively active receptors - i.e. receptors that can activate G proteins without agonist. Such receptors have been widely reported. (1) Is this phenomenon a result of over-expression so that the R<-->R* equilibrium is unaltered but there is more R*, hence more basal activity. Or is it that the equilibrium is actually shifted towards R* in an analogous manner to the presence of agonist. R* is usually stabilized by the binding of both agonist and G protein. If constitutively active receptors only require the G protein, then (2) is it possible that a receptor can exist that only requires agonist to stabilize R*? This would mean that high affinity agonist binding could be obtained without G protein - useful in systems that have no appropriate/ insufficient amounts of G protein. The experiment to show this is relatively simple but I was wondering whether this phenomenon has already been shown. Dan Donnelly Dept Biochem + Mol Biol, University of Leeds, Leeds LS2 9JT, UK. bmb6hdd@biovax.leeds.ac.uk From owner-7tms_r@net.bio.net Wed Mar 29 23:00:00 1995 Path: biosci!rutgers!uwm.edu!msunews!gmi!srvr1.flint.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Date: 30 Mar 1995 15:15:54 GMT Organization: University of Michigan Lines: 86 Message-ID: <3lehva$rm8@lastactionhero.rs.itd.umich.edu> References: <26E101BA_000DF7E0.0098E183748AF7D3$28_1@UK.AC.LEEDS.BIO.VAX> NNTP-Posting-Host: warbler.med.umich.edu BMB6HDD@VAX.BIO.LEEDS.AC.UK wrote: > Constitutively active receptors - i.e. receptors that can activate G > proteins without agonist. > > Such receptors have been widely reported. (1) Is this phenomenon a result of > over-expression so that the R<-->R* equilibrium is unaltered but there is > more R*, hence more basal activity. Or is it that the equilibrium is actually > shifted towards R* in an analogous manner to the presence of agonist. Both phenomena have been pretty clearly demonstrated. If you increase expression levels of wild type (e.g. beta receptor in recent Nature article) you can get increased basal activation of G protein and downstream effector. The newer information relates to mutations in receptors (e.g. adrenergic from Lefkowitz's lab plus rhotopsin in retinitis pigmentosa and the LH and TSH receptor mutations in humans. In these cases you get more basal activation of effector at the same or even lower receptor densities. This clearly shows that the receptors either have a greater amount of R* or the R* is different and has a greater Vmax for activating G protein. The former is the simpler hypothesis and so far seems adequate to explain the data. > > R* is usually stabilized by the binding of both agonist and G protein. > If constitutively active receptors only require the G protein, then > (2) is it possible that a receptor can exist that only requires agonist > to stabilize R*? This would mean that high affinity agonist binding could > be obtained without G protein - useful in systems that have no appropriate/ > insufficient amounts of G protein. The experiment to show this is relatively > simple but I was wondering whether this phenomenon has already been shown. A little recognized implication of early data on BARK is that agonist can change the conformation of receptor to some state (either an R' or the activated R* state). Agonist can stimulate BARK phosphorylation of purified beta receptors without G protein being present. This means that the activating conformational change does not require but is enhanced by G protein as you mentioned. Similarly, the meta II state of rhodopsin is induced by light but it is enhanced by the presence of transducin. If you wanted to use high affinity agonist binding to detect the high affinity state of receptor with no G protein present you would have difficulty. In the absence of a limiting amount of G protein (or some fraction of receptor which is unable to couple to G protein) the receptor would show a single apparent affinity even if some fraction is in the "high affinity" state. This is described in a number of papers but two from my lab are: Neubig, R. R., R. D. Gantzos, and R. S. Brasier. 1985. Agonist and antagonist binding to a2-adrenergic receptors in purified membranes from human platelets: implications of receptor-inhibitory nucleotide binding protein stoichiometry. Mol. Pharmacol 28:475-486. Neubig, R. R. 1994. Membrane organization in G protein mechanisms. FASEB J 8:939-946. Finally, it is interesting to me that so many people are surprised about the idea that receptors can activate effectors in the absence of agonist. Coming from the nicotinic acetylcholine receptor field, I think it is well recognized that receptors can be both spontaneously activated Jackson, M. B. 1984. Spontaneous openings of the acetylcholine receptor channel. Proc. Natl. Acad. Sci. U. S. A. 81:3901-3904. and spontaneously desensitized Boyd, N. D. and J. B. Cohen. 1980. Kinetics of binding of [3H]acetylcholine and [3H]carbamylcholine to Torpedo postsynaptic membranes: slow conformational changes of the cholinergic receptor. Biochemistry 19:5344-5353. Also several groups (including mine) have provided evidence for pre-coupling of 7tms_r's to G proteins without agonist and for spontaneous activation without agonist (e.g. Costa for opiate receptors). The new information is most exciting in my view in the potential that these mutations have as a cause of human disease. Rick Neubig http://www.umich.edu/~rneubig From owner-7tms_r@net.bio.net Thu Mar 30 23:00:00 1995 Path: biosci!daresbury!trane.uninett.no!sunic!sunic.sunet.se!seunet!news2.swip.net!doc.news.pipex.net!pipex!lyra.csx.cam.ac.uk!mole.bio.cam.ac.uk!flh From: flh@mole.bio.cam.ac.uk (Frances Hannan (Zoology)) Newsgroups: bionet.molbio.proteins.7tms_r Subject: pM ligands for GPCRs Date: 31 Mar 1995 14:58:03 GMT Organization: University of Cambridge, England Lines: 12 Message-ID: <3lh59r$r2a@lyra.csx.cam.ac.uk> NNTP-Posting-Host: mole.bio.cam.ac.uk Hi all, Are there any cases of ligands which activate their receptors in the picomolar range ie around 10-12? I have heard that some peptides act at 10-10. This sort of thing is difficult to extract using literature searches so thanks for any help you can give... -- Frances Hannan BILMS, Zoology, Downing St, Cambridge, CB2 3EJ, UK Phone (0223)336663, FAX (0223)461954 flh@mole.bio.cam.ac.uk