From owner-7tms_r@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!RULGCA.LEIDENUNIV.NL!IJZERMAN From: IJZERMAN@RULGCA.LEIDENUNIV.NL Newsgroups: bionet.molbio.proteins.7tms_r Subject: school on medicinal chemistry Date: 3 Jul 1995 03:20:11 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 178 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HSFLT0DDXE000Q8Y@rulgca.LeidenUniv.nl> There are a few places left in the IVth School on Medicinal Chemistry (see below for full information). The school is primarily meant for research chemists in pharmaceutical industry wishing to update their knowledge in pharmacology, toxicology and new trends in drug design. If you are interested, please send a fax to Mrs. F.J. Velthorst LACDR PO Box 9502 2300RA Leiden The Netherlands fax 31-71-274277 to register or for further information 24-27 October 1995, Noordwijkerhout, The Netherlands IVth LACDR SCHOOL ON MEDICINAL CHEMISTRY COURSE OUTLINE The development of new drugs has become largely dependent on a deeper understanding of human (patho)physiology. Nowadays, pharmacology and toxicology, both in vitro and in vivo, are essential to exploit fundamental knowledge for the development of drug candidates. As a consequence, both disciplines are more and more applied at an early stage of drug design to guide synthetic strategies. The course comprises basic and advanced aspects of lead finding, pharmacology and toxicology to provide research chemists in the pharmaceutical industry with the appropriate background for their daily practice. Topics include the impact of new research fields and techniques, such as molecular biology and molecular modelling, on drug research, as well as a thorough introduction in pharmacodynamics, pharmacokinetics and molecular toxicology. New in the 4th School will be the topic of COMBINATORIAL CHEMISTRY and molecular diversity. In addition, three case histories will give a flavour of chance and strategy in drug development. Essential to the course is a workshop-like case-study ('the design of a new drug') in which all participants are actively engaged. Acclaims from participants to the previous schools: excellent documentation...; case-study very useful....; very clear review of drug research today....; lectures very good....; beyond expectation....; intellectually most stimulating. COURSE DIRECTION Prof.dr. H. Timmerman, Director of Research Leiden/Amsterdam Center for Drug Research Head Department of Pharmacochemistry, Faculty of Chemistry, Vrije Universiteit Amsterdam, The Netherlands. His research programme mainly concerns the different histamine receptors and their ligands. He is a member of the editorial boards of several international journals and editor of the series Pharmaco- chemistry Library (Elsevier) and Methods and Principles in Medicinal Chemistry (VCH). Dr. A.P. IJzerman Associate Professor, Division of Medicinal Chemistry, Leiden- /Amsterdam Center for Drug Research, Leiden University, The Netherlands. Dr. IJzerman supervises the receptor research group at the Center, and is involved in research on purinergic receptors and drug design. He has ample experience in modern techniques to study ligand-receptor interactions, such as molecular model- ling/computer graphics. He is a referee to major international journals in pharmacology, medicinal chemistry and pharmaceutics. REGISTRATION INFORMATION The course will be held at the Leeuwenhorst Congress Center, Noordwijkerhout, The Netherlands. Tel. 31 2523 77106. Hotel accommodation: A number of fully equipped single rooms have been reserved at the Congress Center. Fee Individuals: Hfl. 2.500,-. This includes course documentation, mid-session refreshments, lunches, dinners and hotel accommodation with breakfast (3 nights). Group fee: Hfl. 2.500,- for the first participant and Hfl. 2.250,- for the second and following participants from the same organization. Ph.D. students: Up to five Ph.D. students may attend the course for a reduced fee. A statement of the supervisor acknowledging the status of Ph.D. student should accompany the registration form. Registration and payment: It is advised to forward the registration form as soon as possible in view of the limited course capacity of 25 participants. Confirmation of registration will be returned upon receipt, together with an invoice for the course fee. Registration will not be final until payment is received. The organization reserves the right to cancel the course should the number of registrations be lower than 12. Notice of cancellation, with a full refund, will be given before September 1995. Cancellations: Cancellations with a full refund may be made until September 1, 1995. Cancellations between September 1 and October 1, 1995: 50% refund. No refund is possible on cancellations received after this date. Substitutions may be made at any time. The official language of the course is English. COURSE PROGRAMME Tuesday, October 24, 9.30 - 10.00 Registration Pharmacology 10.00 - 11.00 Mathematics of receptor-ligand interaction dr. A.P. IJzerman, Leiden, NL 11.15 - 12.15 Receptor binding in industrial perspective dr. M. Tulp, Weesp, NL 13.30 - 14.30 Signal transduction and second messengers prof.dr. A. Bast, Amsterdam NL 14.30 - 15.30 Selection of pharmacological models dr. C.L.E. Broekkamp, Oss, NL 16.00 - 17.00 Molecular biology in drug design dr. R. Leurs, Amsterdam NL 20.30 - 21.30 Case history 'anti-migraine agents' dr. F.P. van de Wijngaert, Zeist, NL Wednesday, October 25 Pharmacology (cont). 9.00 - 10.15 Clinical pharmacology: what happens to your molecule? prof.dr. A.F. Cohen, Leiden, NL 10.45 - 11.30 Case study in drug design: introduction dr. N. de Hoog, Haarlem, NL 11.30 - 12.30 The concept of drug selectivity prof.dr. H. Timmerman, Amsterdam,NL 13.30 - 17.30 Case study in drug design participants 20.30 - 21.30 Case history 'Omeprazole' dr. P. Lindberg, Molndal, Sweden Thursday, October 26 Fate of drugs 9.00 - 10.30 Introduction to the pharmacokinetics of drugs dr. W. Meuldermans, Beerse, B 11.00 - 12.00 Toxicology in industry dr. W. Coussement, Beerse, B 12.00 - 13.00 In vitro toxicology in drug research prof.dr. G.J. Mulder, Leiden, NL Lead finding and optimization 14.30 - 15.30 Combinatorial chemistry prof.dr. H.C.J. Ottenheijm, Oss, NL 16.00 - 17.00 Screening of biological materials dr. C.J. Latham, Slough, GB 20.30 - 21.30 Case history 'neuromuscular blockers' dr. L. van den Broek, Oss, NL Friday, October 27 Lead finding and optimization (cont.) 9.00 - 10.00 3D-databases in drug design dr. S. van Helden, Oss, NL 10.30 - 11.30 Molecular modeling in drug design prof.dr. J. Tollenaere, Beerse, B 11.30 - 12.30 Novel targets for drug design prof.dr. F. Nijkamp, Utrecht, NL 14.00 - 16.00 Case study in drug design: presentation participants REGISTRATION FORM IVth LACDR School on Medicinal Chemistry 24-27 October 1995 Name: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Organization: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Position: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Address:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Postal Code:. . . . . . . . . . . . . . . . . . . . . . . . City: . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . State/country:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Telephone:. . . . . . . . . . . . . . . . . . . . . . . . . Telefax:. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Date of arrival: Date of Departure: Signature: Return by fax or regular mail to: LACDR-Secretariat, c/o Mrs. F.J. Velthorst, P.O. Box 9502, 2300 RA Leiden, The Netherlands Phone 31 71 274341; Fax 31 71 274277 From owner-7tms_r@net.bio.net Sun Jul 02 23:00:00 1995 Path: biosci!chemie.uni-regensburg.de!Stefan.Dove From: Stefan.Dove@chemie.uni-regensburg.de ("Stefan Dove ", t 4673) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor States Date: 3 Jul 1995 05:41:05 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 51 Sender: daemon@net.bio.net Distribution: world Message-ID: <100BF0F8436B@alf5.ngate.uni-regensburg.de> From my viewpoint of a molecular modeler, the discussion about activated and non-activated receptor states is, although useful, somewhat too simplifying. We sometimes forget, that we have to do with dynamic equilibria of a lot of conformational states (not only of two, four, or six), whose probabilities result from statistical thermodynamics (Maxwell-Boltzmann distribution). E. g., a snapshot of the interacting, energy-minimized seven TM helices can never explain how the ligands approach their "final" binding sites. In my imagination, the helices do oscillate along different directions, and the ligand molecules are transported by "gliding" along dynamic potential surfaces. A lot of transient local minima states and energy barriers are passed. The "final" ligand-receptor complex is energetically more favorable and insofar more frozen. Then some degrees of freedom of helical movements are reduced, and others eventually favored. This is not contradictory to the model of Zhang and Weinstein (J. Med. Chem. 36, 934 - 938 (1993)), who suggested from molecular dynamics simulations especially large movements of helices 5 and 6 with bound full agonists, but not with antagonists. Of course, the resulting conformational change, accompanied by stabilized helical structures, cannot be simulated without consideration of the cytoplasmatic loop 3 (which is not possible up to now and will depend on refined crystal structures of G-protein coupled receptors). Assuming a distribution of a lot of interconverting conformations also for C 3, neither R-G procoupling nor constitutively active receptors (RG - RG* equilibrium) are surprising. But obviously a definite change of this region is requested for strongly stabilizing G-protein bound states. This change could be more likely (more frequently) induced by larger movements of helices 5 and 6. The "high affinity" state of agonist binding needs a stable ARG* complex. That means that the conformational change of the C 3 region is frozen only with G-protein bound. Then, oszillations of helices 5 and 6 are restricted, and agonist binding is further stabilized. Such simple and rather mechanistic considerations do not significantly expand our understanding about ligand-receptor-effector interactions, but are helpful in coming from static to more dynamic models. It should be noted, that all hitherto existing mathematical formulations of ternary complex models are approximations to "multi-state" models, as they only neglect the huge multiplicity of states of one, two, or three "reactants". The complexity of the binding equations only depends on the number of the reactants (A, R, and G). Insofar, my considerations are more philosophic than of practical relevance with respect to the analysis of binding curves of agonists (negative cooperativity). Stefan Dove, Univ. Regensburg, Inst. Pharmacy From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!DEFIANCE.HSC.COLORADO.EDU!port_d From: port_d@DEFIANCE.HSC.COLORADO.EDU ("Dave Port") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang II receptors Date: 5 Jul 1995 07:25:22 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507031604.KAA12386@essex.hsc.colorado.edu> Ang II receptors Does anybody know of a cell line "confirmed" to not express AT1/2 receptors? I'm guessing CHO-K1's will be ok, but I don't know this for a fact. Any information would be appreciated. Thanks, J.D. Port UCHSC MED/PHARM port_d@defiance.hsc.colorado.edu From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!news.sprintlink.net!europa.chnt.gtegsc.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: On the meaning of R and R* Date: 4 Jul 1995 10:40:42 GMT Organization: University of Michigan Lines: 34 Message-ID: <3tb5ra$sv8@lastactionhero.rs.itd.umich.edu> References: <3ss7io$l6b@gaia.ucs.orst.edu> <3sv77s$8j0@maureen.teleport.com> 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) Kim Neve wrote: >vogelw@ava.bcc.orst.edu (Walter Vogel) wrote: >> The essential advance of this >>model (also found in ref. 3) is that there is an isomerization of the >>receptor from R to R* and that only R* is capable of interacting with G >>proteins to form R*G. >> > >This point was also made explicitly by Contreras and Molinoff in two >papers published in 1986 (JPET 237:165-172, and JPET 239:136-143). >Interactions of Beta-2 receptors with G proteins were prevented by GTP in >one paper and by solubilization in the second paper, yet both still >provided evidence for an agonist-dependent conformational change from R >to R*. > >Kim Neve >Oregon Health Sciences University > Actually, strong evidence for the existence of an R <> R* equilibrium (or at least an ability of agonists to cause a conformational change independent of RG coupling) was present in Lefkowitz's data that agonists stimulated the BARK-dependent phosphorylation of purified beta adrenergic receptors!. Benovic-JL; Kuhn-H; Weyand-I; Codina-J; Caron-MG; Lefkowitz-RJ Proc-Natl-Acad-Sci-U-S-A. 1987 Dec; 84(24): 8879-82 _________________________________________________________ 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 Jul 04 23:00:00 1995 Path: biosci!DEFIANCE.HSC.COLORADO.EDU!port_d From: port_d@DEFIANCE.HSC.COLORADO.EDU ("Dave Port") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang II receptors Date: 5 Jul 1995 07:20:08 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507032016.OAA22183@essex.hsc.colorado.edu> Ang II receptors Does anybody know of a cell line "confirmed" to not express AT1/2 receptors? I'm guessing CHO-K1's will be ok, but I don't know this for a fact. Any information would be appreciated. Thanks, J.D. Port UCHSC MED/PHARM port_d@defiance.hsc.colorado.edu From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!DEFIANCE.HSC.COLORADO.EDU!port_d From: port_d@DEFIANCE.HSC.COLORADO.EDU ("Dave Port") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang II receptors Date: 5 Jul 1995 07:20:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507041620.KAA06770@essex.hsc.colorado.edu> Ang II receptors Does anybody know of a cell line "confirmed" to not express AT1/2 receptors? I'm guessing CHO-K1's will be ok, but I don't know this for a fact. Any information would be appreciated. Thanks, J.D. Port UCHSC MED/PHARM port_d@defiance.hsc.colorado.edu From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!CHEM.VU.NL!beukers From: beukers@CHEM.VU.NL (Margot Beukers) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Subscription Date: 5 Jul 1995 07:00:06 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <9507051355.AA09544@gallium.chem.vu.nl> In mailing this I would like to ask for a subscription to your network. Thanks in advance, Margot Beukers From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!news.sprintlink.net!europa.chnt.gtegsc.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Receptor States Date: 4 Jul 1995 11:00:36 GMT Organization: University of Michigan Lines: 35 Message-ID: <3tb70k$sv8@lastactionhero.rs.itd.umich.edu> References: <100BF0F8436B@alf5.ngate.uni-regensburg.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) To: Stefan.Dove@chemie.uni-regensburg.de Stefan.Dove@chemie.uni-regensburg.de ("Stefan Dove ", t 4673) wrote: >From my viewpoint of a molecular modeler, the discussion about >activated and non-activated receptor states is, although useful, >somewhat too simplifying. We sometimes forget, that we have to do >with dynamic equilibria of a lot of conformational states (not only >of two, four, or six), whose probabilities result from statistical >thermodynamics (Maxwell-Boltzmann distribution). Stefan, You are absolutely right that the number of receptor states is probably much greater than we experimentalists can define but we do need to start somewhere. Once you start putting in the receptor and G protein states plus the kinetic parameters for the transitions between those states, the models very rapidly outstrip what the data can define. You might be interested in the discussion in my minireview article in BioEssays 11:136-140, 1989 entitled "How does a key fit a flexible lock? Structure and dynamics in receptor function". Although I didn't have the detailed modelling information that you cite, I pointed out that we need to consider multiple states of receptors. Thus even the classical allosteric enzyme Aspartate Transcarbamylase has been shown to have not two but multiple different conformations. The nictotinic acetylcholine receptor (where my early scientific background was) has multiple states (at least 4 - resting, active, and 2 desensitized). We shouldn't be surprised if the same is true of the 7tmr's, once we get powerful enough tools to look at them that carefully. Rick _________________________________________________________ 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 Jul 04 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!news.sprintlink.net!cam.news.pipex.net!pipex!dish.news.pipex.net!pipex!oleane!jussieu.fr!univ-lyon1.fr!serra.unipi.it!usenet From: Roberto Buccione Newsgroups: bionet.molbio.proteins.7tms_r Subject: Intracellular 7TMD receptors? Date: 4 Jul 1995 15:36:29 GMT Organization: Consorzio Mario Negri Sud, ITALY Lines: 10 Message-ID: <3tbn5t$g1i@serra.unipi.it> NNTP-Posting-Host: neurobio03.cmns.mnegri.it 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 Is there strong evidence showing an intracellular localization of 7tm receptors other than on the PM, besides when they are overexpressed? And anyway, what would be the meaning of this? In any case, heterotrimeric G proteins happen to be all over the cell, certainly not only on the PM... We would like to discuss this, especially in the light of possible homeostatic functions they might have in intracellular regulation (metabolism, cytoskeleton, etc.). From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 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: Re: secretin GPCR family mutants etc Date: 5 Jul 1995 08:43:58 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: <9507051842.AA6825@pho018.sb.com> NNTP-Posting-Host: net.bio.net A technical question to add to the information requested by Dan Donnelly. Concerning the secretin class of receptors, what expression systems have been used for over expression and or signal transduction studies? Are there any subclasses of G-proteins that are preferentially utilized by these members of this subfamily? 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 From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!IUNHAW1.IUN.INDIANA.EDU!WANDERS From: WANDERS@IUNHAW1.IUN.INDIANA.EDU ("W. Marshall Anderson") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Substance P antagonist Date: 5 Jul 1995 09:42:20 -0700 Organization: Indiana University Northwest Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: Reply-To: wanders@iunhaw1.iun.indiana.edu NNTP-Posting-Host: net.bio.net What is currently considered to be the best substance P antagonist (with special reference to studies of the eye if possible?) Reply to 7tms_r@net.bio.net or wanders@iunhaw1.iun.indiana.edu and I will post the responses to the group. Thanks, W. Marshall Anderson I.U. Sch. Med. NWCME 3400 Broadway Gary, IN 46408 USA phone 219 n980-6534 FAX 219 980-6566 e-mail wanders@iunhaw1.iun.indiana.edu W. Marshall Anderson, Ph.D. Phone: 219 980-6534 FAX: 219 980-6566 E-mail: wanders@iunhaw1.iun.indiana.edu From owner-7tms_r@net.bio.net Tue Jul 04 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!Dortmund.Germany.EU.net!nntp.gmd.de!news.rwth-aachen.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!news.belwue.de!fu-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: On the meaning of R and R* Date: 5 Jul 1995 18:44:56 GMT Organization: University of Wuerzburg, Germany Lines: 51 Message-ID: <3temj8$g67@winx03.informatik.uni-wuerzburg.de> References: <3ss7io$l6b@gaia.ucs.orst.edu> <3sv77s$8j0@maureen.teleport.com> <3tb5ra$sv8@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: > Kim Neve wrote: > >vogelw@ava.bcc.orst.edu (Walter Vogel) wrote: > >> The essential advance of this > >>model (also found in ref. 3) is that there is an isomerization of the > >>receptor from R to R* and that only R* is capable of interacting with G > >>proteins to form R*G. > > > >This point was also made explicitly by Contreras and Molinoff in two > >papers published in 1986 (JPET 237:165-172, and JPET 239:136-143). > >Interactions of Beta-2 receptors with G proteins were prevented by GTP in > >one paper and by solubilization in the second paper, yet both still > >provided evidence for an agonist-dependent conformational change from R > >to R*. > Actually, strong evidence for the existence of an R <> R* equilibrium > (or at least > an ability of agonists to cause a conformational change independent of RG > coupling) was present in Lefkowitz's data that agonists stimulated the > BARK-dependent phosphorylation of purified beta adrenergic receptors. > Benovic-JL; Kuhn-H; Weyand-I; Codina-J; Caron-MG; Lefkowitz-RJ > Proc-Natl-Acad-Sci-U-S-A. 1987 Dec; 84(24): 8879-82 As far as I understand this issue it has not been demonstrated yet that the R* to which the receptor kinase binds is different from the R* to which the G protein binds. (I am not sure how this relates to the original question :-) However, there are hints that the binding sites for the two proteins on the receptor are different: @article{benovic:90, author = {J. L. Benovic and J. Onorato and M. J. Lohse and H. G. Dohlman and Claudia Staniszewski and M. G. Caron and R. J. Lefkowitz}, title = {Synthetic peptides of the hamster $\beta_2$-adrenoceptor as substrates and inhibitors of the $\beta$-adrenoceptor kinase.}, journal = {Br. J. Clin. Pharmacol.}, volume = 30, pages = {3S--12S}, year = 1990 } --Cornelius. -- /* 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 Wed Jul 05 23:00:00 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: Re: secretin GPCR family mutants etc Date: 6 Jul 1995 08:58:00 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 19 Sender: daemon@net.bio.net Distribution: world Message-ID: <9507061722.AA1316@pho018.sb.com> NNTP-Posting-Host: net.bio.net A technical question to add to the information requested by Dan Donnelly. Concerning the secretin class of receptors, what expression systems have been used for over expression and or signal transduction studies? Are there any subclasses of G-proteins that are preferentially utilized by these members of this subfamily? Jonathan A. Lee, Ph.D. 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 From owner-7tms_r@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!BIOMED.MED.YALE.EDU!KARNE From: KARNE@BIOMED.MED.YALE.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 7 Jul 1995 14:14:09 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HSLHAGVE0I005U4L@BIOMED.MED.YALE.EDU> NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!NAXOS.UNICE.FR!mazella From: mazella@NAXOS.UNICE.FR (mazella) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ang II receptors Date: 7 Jul 1995 09:21:41 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Ang II receptors Does anybody know of a cell line "confirmed" to not express AT1/2 receptors? I'm guessing CHO-K1's will be ok, but I don't know this for a fact. Any information would be appreciated. Thanks, J.D. Port UCHSC MED/PHARM port_d@defiance.hsc.colorado.edu From owner-7tms_r@net.bio.net Thu Jul 06 23:00:00 1995 Path: biosci!rutgers!uwm.edu!cs.utexas.edu!news.sprintlink.net!europa.chnt.gtegsc.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Ang II receptors Date: 7 Jul 1995 12:21:10 GMT Organization: University of Michigan Lines: 14 Message-ID: <3tj8rm$48c@lastactionhero.rs.itd.umich.edu> References: <199507031604.KAA12386@essex.hsc.colorado.edu> NNTP-Posting-Host: pm047-23.dialip.mich.net Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) To: oluwole@umich.edu We're expressing rat AT1a receptors in HEK293 cells and there is no significant binding of 125I-AII and no detectable AII-stimulated IPx release in the cells prior to transfection. We also did some pilots in COS cells (I believe COS1 but it could be COS7) and didn't see any binding or reponses there either. See our abstract in FASEB J this spring. -- Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 From owner-7tms_r@net.bio.net Fri Jul 07 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!nntp.gmd.de!news.rwth-aachen.de!news.rhrz.uni-bonn.de!news.uni-stuttgart.de!news.belwue.de!fu-berlin.de!cs.tu-berlin.de!fauern!winx03!wrzx15!phak004 From: phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Sigler talk Date: 7 Jul 1995 15:19:10 GMT Organization: University of Wuerzburg, Germany Lines: 30 Message-ID: <3tjj9e$hra@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] I just come back from a talk given by Paul Sigler and want to give a summary, as promised. Sigler talked almost exclusively on the structure of the alpha subunit of transducin and not, as promised, on the structure of the holotrimer. I didn't write much down since it appeared to me that he related mainly material that is already published. Frankly, I was quite disappointed (I have never understood the fascination of structural biochemists with reaction mechanisms :-). He did give some hints on the structure of the heterotrimer. They use a chimera between transducin-alpha and the N-terminus of Gi-alpha. The heterotrimer is not acylated. He did not mention the expression system used. The crystals apparently diffract to high resolution. They have been able to refine the structure of the beta-subunit so far (not those of the gamma-subunit). The N-terminus of the alpha subunit is well- defined. At the moment, Paul Sigler has a collaboration with Volker Hildebrandt from Munich where they try to investigate interactions between transducin and rhodopsin. That's it, folks. Sorry :-/ --Cornelius. -- /* 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 Sun Jul 09 23:00:00 1995 Path: biosci!rutgers!uwm.edu!news.moneng.mei.com!howland.reston.ans.net!news.sprintlink.net!uunet!in1.uu.net!emi.com!pauling.wadsworth.org!usenet From: Meg Davis-Cox Newsgroups: bionet.molbio.proteins.7tms_r Subject: cGMP dependent protein kinase Date: 10 Jul 1995 20:29:37 GMT Organization: Wadsworth Center Lines: 3 Message-ID: <3ts2jh$ape@pauling.wadsworth.org> NNTP-Posting-Host: alcor.wadsworth.org Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) I am looking for a source of cGMP dependent protein kinase (purified or recombinant). Does anyone have any ideas? Thanks! From owner-7tms_r@net.bio.net Sun Jul 09 23:00:00 1995 Path: biosci!MAIL.MED.CORNELL.EDU!egraaka From: egraaka@MAIL.MED.CORNELL.EDU (liz raaka) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Subscribe Date: 10 Jul 1995 14:44:37 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 2 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507102142.RAA25329@mail.med.cornell.edu> NNTP-Posting-Host: net.bio.net SUBSCRIBE. From owner-7tms_r@net.bio.net Mon Jul 10 23:00:00 1995 Path: biosci!RISC.IDG.FI.CNR.IT!giolitti From: giolitti@RISC.IDG.FI.CNR.IT (Alessandro Giolitti) Newsgroups: bionet.molbio.proteins.7tms_r Subject: B1 bradykinin receptor sequence Date: 11 Jul 1995 03:22:04 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 26 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hi all: has (as I expect) the B1 bradykinin receptor been cloned? I need the sequence, that should have been published somewhere (but I have no papers about that). Thanks for any info, Alessandro Giolitti _______________________________________ Dr.Alessandro Giolitti Drug Design Dept. Menarini s.r.l. Via Sette Santi, 3 50131 Firenze Italy Ph. : +39-55-5680240 FAX : +39-55-5680419 e-mail: giolitti@risc.idg.fi.cnr.it _______________________________________ From owner-7tms_r@net.bio.net Tue Jul 11 23:00:00 1995 Path: biosci!MED.UNC.EDU!erweiss From: erweiss@MED.UNC.EDU (Ellen R. Weiss) Newsgroups: bionet.molbio.proteins.7tms_r Subject: re:phosphorylation Date: 12 Jul 1995 08:54:10 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: <9507121552.AA13061@garner.med.unc.edu> NNTP-Posting-Host: net.bio.net Dave Port (port_d@DEFIANCE.HSC.COLORADO.EDU) wrote: > Is anyone out there aware of any information on hierarchical > phosphorylation of any G-protein coupled receptor? You should also look at: Prossnitz, E. R., Kim, C. M., Benovic, J. L., & Ye, R. D. (1995). Phosphorylation of the n-formyl peptide receptor carboxyl terminus by the G protein-coupled receptor kinase, GRK2. J Biol Chem, 270(3), 1130-1137. They demonstrate hierarchical phosphorylation by mutagenesis using Glu substitutions. Ellen Weiss University of North Carolina From owner-7tms_r@net.bio.net Thu Jul 13 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!plug.news.pipex.net!pipex!tank.news.pipex.net!pipex!sunsite.doc.ic.ac.uk!news.cc.ic.ac.uk!usenet From: Mark Pallen Newsgroups: bionet.molbio.proteins.7tms_r Subject: disulphide bridges Date: 14 Jul 1995 15:35:10 GMT Organization: Imperial College Lines: 27 Message-ID: <3u62re$jnu@oban.cc.ic.ac.uk> NNTP-Posting-Host: mpm1.bc.ic.ac.uk 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 Hi! A colleague of mine, Gadi Frankel (gfrankel@molbiol.ox.ac.uk) has asked me to post the following request: If you have a protein with two or more cyteines in it, what is the best and/or easiest biochemical method for determining which, if any, pair or pairs of cysteines is/are linked via a disulphide bridge? please e-mail as well as post. thanks in advance Mark ******************************************************** Dr Mark Pallen, Senior Lecturer in Medical Microbiology, St Bartholomew's Hospital Medical College, London, EC1A 7BE currently Visiting Scientist at Imperial College Rm 502, Dept of Biochem, Imperial College, London, SW7 2AY email:m.pallen@ic.ac.uk WWW: http://www.qmw.ac.uk/~rhbm001/mpallen.html phone: day ++44(0)1715945254, eves ++44(0)1815057937, FAX ++44(0)1715945255 ******************************************************** "My country is the world and my religion is to do good..." Thomas Paine ******************************************************** From owner-7tms_r@net.bio.net Thu Jul 13 23:00:00 1995 Path: biosci!DNAX.ORG!nancy_fadis From: nancy_fadis@DNAX.ORG ("Nancy Fadis") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Sept. California Meeting Date: 14 Jul 1995 12:42:37 -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 G-Protein Coupled Receptors Group: Are any of you aware of a meeting on either G-coupled receptors or receptor-ligand interactions that will take place in the San Diego, California area in September of this year? Or anything even close? Nancy Fadis DNAX Research Institute Palo Alto, CA nancy_fadis@dnax.org From owner-7tms_r@net.bio.net Sun Jul 16 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU (Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D.") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Radioligand Binding Technology Date: 17 Jul 1995 10:30:34 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 38 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507171728.NAA03319@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > To those who bind, > > I am curious about current preferences in binding technology. For small > assays we have been using the Millipore bucket-thing and plenty of people > around here have Brandel harvesters. I am considering buying a gizmo for > moderate sized assays. I am interested, and perhaps others are in thoughts > regarding ease of use, reliability, consistency, rinsing uniformity, > speed, clogging problems (all the important minutiae) of the currently > available gadgets. > Ted: Sorry for the tardy reply, but I have been in grant mode and symposium mode and are only now returning to email mode. I go with Brandel, and I would recommend Teflon tubing for most applications. It speeds flow and without it there can be significant trapping of lipophilic drugs and membranes in the Tygon tubing. Be certain to get a large enough pump. Compare pumps by liters/min flow rate, not by horsepower or the ultimate vacuum. Use the shortest and widest vacuum tubing possible. If you insist upon using massive harvesters, remember that you need 8 times the flow rate with a 96-well than with a 12-well, because you are dividing your liters/min air flow into 1/96th part. The new membrane or oilless pumps are quite nice. Other fine points: use the thinnest filter you can get away with in terms of protein capacity, as this will reduce nonspecific binding. We use GF/C for most things (up to 200 ug protein/well) and GF/B equivalent for higher protein. S & S will custom-make filters to fit Brandel harvesters --they offer more variety and lower cost. Compare filter pre-treatment protocols-- we use higher PEI for substance P (0.3%) compared to other receptors (0.1%) and for angiotensin II we pretreat with BSA becuase PEI actually increases nonspecific binding. PEI solution MUST be cold when contacting the filter. We go overnight in most cases (a must for substance P) or at least 4h. If you soak warm, PEI will dossolve the filter (pH is around 9). I hope you find this lab lore useful. From owner-7tms_r@net.bio.net Sun Jul 16 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU (Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D.") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: intracellular 7TM receptors? Date: 17 Jul 1995 10:38:05 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 28 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507171736.NAA03540@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > Is there strong evidence showing an intracellular localization of 7tm > receptors other than on the PM, (besides when they are overexpressed)? > And anyway, what would be the meaning of this? > In any case, heterotrimeric G proteins happen to be all over the cell, > certainly not only on the PM... > We would like to discuss this, especially in the light of possible > homeostatic functions they might have in intracellular regulation > (metabolism, cytoskeleton, etc.). > Setting aside the question of mitochondrial receptors, many garden-variety 7tms_r are predominantly located intracellularly. This was shown at Cornell a few years back by Virginia Pickel and Chiye Aoki (now at NYU) who did electron microscopy of beta1 and beta2 adrenergic receptors and found a primarily intracellular localization. Many were associated with the golgi and were presumably undergoin transport and posttranslational modification, but others were in vesicular structures (internalized?). A minority were actually embedded in the plasma membrane. There are several caveats, such as the effects of fixation, and the possibly reduced antigenicity of a PM-embedded and tightly folded protein versus a possibly more extended conformation in other compartments. But it seems clear that beta-receptors spend a good portion of their life cycle away from the PM. This also agrees with subcellular fractionation data, by the way, but we all dismissed "intracellular receptors" as an artifact of the fractionation method. From owner-7tms_r@net.bio.net Mon Jul 17 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!swrinde!dish.news.pipex.net!pipex!oleane!jussieu.fr!univ-lyon1.fr!swidir.switch.ch!scsing.switch.ch!news.rediris.es!acebo.sdi.uam.es!samba.cnb.uam.es!jcastilla From: jcastilla@samba.cnb.uam.es Newsgroups: bionet.molbio.proteins.7tms_r Subject: Looking for antibodies Date: 18 Jul 95 10:40:27 WET Organization: C.N.Biotecnologia, CSIC Lines: 6 Message-ID: <1995Jul18.104027.1@samba.cnb.uam.es> NNTP-Posting-Host: samba.cnb.uam.es We are very interested in getting some information about antibodies that recognize mouse J chain and porcine secretory component (and companies that commercialize them or laboratory that use them). Thank you very much. From owner-7tms_r@net.bio.net Mon Jul 17 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news.sprintlink.net!tank.news.pipex.net!pipex!oleane!jussieu.fr!univ-lyon1.fr!swidir.switch.ch!scsing.switch.ch!news.belwue.de!fu-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: intracellular 7TM receptors? Date: 18 Jul 1995 11:13:06 GMT Organization: University of Wuerzburg, Germany Lines: 29 Distribution: world Message-ID: <3ug502$hnk@winx03.informatik.uni-wuerzburg.de> References: <199507171736.NAA03540@slc5.INS.CWRU.Edu> NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D. (pre@PO.CWRU.EDU) wrote: > Setting aside the question of mitochondrial receptors, many garden-variety > 7tms_r are predominantly located intracellularly. This was shown at > Cornell a few years back by Virginia Pickel and Chiye Aoki (now at NYU) > who did electron microscopy of beta1 and beta2 adrenergic receptors and > found a primarily intracellular localization. Many were associated with > the golgi and were presumably undergoin transport and posttranslational > modification, but others were in vesicular structures (internalized?). If there exists a paper about these observations, could you please post the reference? I am aware of several electron microscopy studies on the beta2-AR from the labs of Donny Strosberg and Kai Simons, but I have never heard of the persons that you have mentioned -- therefore I would be very interested in any pictures :-) Originally it was believed that internalized beta2-ARs could not signal any more and thus internalization was believed to play an important role in desensitization. This view has recently been challenged. Internalization may take place in caveolae, where at least some G proteins are enriched; thus signalling of the internalized receptor has become a possibility. In addition, it seems that internalization leads to resensitization (i.e. removal of arrestins and phosphate groups) of receptors. --Cornelius. -- /* 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 Mon Jul 17 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU (Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D.") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Ang II receptors Date: 18 Jul 1995 07:43:09 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507181441.KAA12262@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > Ang II receptors > Does anybody know of a cell line "confirmed" to not express AT1/2 receptors? > I'm guessing CHO-K1's will be ok, but I don't know this for a fact. Any > information would be appreciated. > Many lines of CHO and COS cells express Ang IV receptors, which can become labeled in 125I-Ang II binding assays if the ligand degrades. Inclusion of phosphoramidon along with PMSF, leupeptin, and bacitracin should prevent formation of Ang IV. From owner-7tms_r@net.bio.net Mon Jul 17 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU (Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D.") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Substance P antagonist Date: 18 Jul 1995 07:39:13 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507181437.KAA12111@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > What is currently considered to be the best substance P antagonist (with > special reference to studies of the eye if possible?) > CP 99,994 is the most potent non-peptide NK-1 antagonist. There is more literature on CP 96,345 but it is a little weaker and has calcium channel blocking activity. It is available from Pfizer in Groton, CT. From owner-7tms_r@net.bio.net Tue Jul 18 23:00:00 1995 Path: biosci!rutgers!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 TOCs for 7tms_r Date: 19 Jul 1995 12:05:41 GMT Organization: Mass. General Hospital, Renal Unit Lines: 389 Distribution: world Message-ID: NNTP-Posting-Host: receptor.mgh.harvard.edu Here is a large block of these 6/9-8/4. I hope I did not miss any! AU Borkowski-J-A. Ransom-R-W. Seabrook-G-R. Trumbauer-M. Chen-H. Hill-R-G. Strader-C-D. Hess-J-F. TI Targeted disruption of a B2 bradykinin receptor gene in mice eliminates bradykinin action in smooth muscle and neurons SO J-Biol-Chem. 1995 June 9. 270(23). p 13706. AU Bouaboula-M. Bourrie-B. Rinaldi-Carmona-M. Shire-D. Fur-G-L. Casellas-P. TI Stimulation of cannabinoid receptor CB1 induces krox-24 expression in human astrocytoma cells SO J-Biol-Chem. 1995 June 9. 270(23). p 13973. AU Kim-J. Wess-J. Rhee-A-M-. Schoneberg-T. Jacobson-K-A. TI Site-directed mutagenesis identifies residues involved in ligand recognition in the human A2a adenosine receptor SO J-Biol-Chem. 1995 June 9. 270(23). p 13987. AU Yamano-Y. Ohyama-K. Kikyo-M. Sano-T. Nakagomi-Y. Inoue-Y. Nakamura-N. Morishima-I. Guo-D-F. Hamakubo-T. Inagami-T. TI Mutagenesis and the molecular modeling of the rat angiotensin II receptor (AT1) SO J-Biol-Chem. 1995 June 9. 270(23). p 14024. AU Chen-H. Kendall-D-A. TI Artificial transmembrane segments. Requirements for stop transfer and polypeptide orientation. SO J-Biol-Chem. 1995 June 9. 270(23). p 14115. AU Pang-J-H-S. Hung-R-Y. Wu-C-J. Fang-Y-Y. Chau-L-Y. TI Functional characterization of the promoter region of the platelet-activating factor receptor gene. Identification of an initiator element essential for gene expression in myeloid cells. SO J-Biol-Chem. 1995 June 9. 270(23). p 14123. AU Pumiglia-K-M. LeVine-H. Haske-T. Habib-T. Jove-R. Decker-S-J. TI A direct interaction between G-protein {beta}{gamma} subunits and the Raf-1 protein kinase SO J-Biol-Chem. 1995 June 16. 270(24). p 14251. AU Ohguro-H. Hooser-J-P-V. Milam-A-H. Palczewski-K. TI Rhodopsin phosphorylation and dephosphorylation in vivo SO J-Biol-Chem. 1995 June 16. 270(24). p 14259. AU Slepak-V-Z. Artemyev-N-O. Zhu-Y. Dumke-C-L. Sabacan-L. Sondek-J. Hamm-H-E. Bownds-M-D. Arshavsky-V-Y. TI An effector site that stimulates G-protein GTPase in photoreceptors SO J-Biol-Chem. 1995 June 16. 270(24). p 14319. AU Rathouz-M-M. Vijayaraghavan-S. Berg-D-K. TI Acetylcholine differentially affects intracellular calcium via nicotinic and muscarinic receptors on the same population of neurons SO J-Biol-Chem. 1995 June 16. 270(24). p 14366. AU Holtmann-M-H. Hadac-E-M. Miller-L-J. TI Critical contributions of amino-terminal extracellular domains in agonist binding and activation of secretin and vasoactive intestinal polypeptide receptors. Studies of chimeric receptors. SO J-Biol-Chem. 1995 June 16. 270(24). p 14394. AU Abrams-C-S. Wu-H. Zhao-W. Belmonte-E. White-D. Brass-L-F. TI Pleckstrin inhibits phosphoinositide hydrolysis initiated by G-protein-coupled and growth factor receptors. A role for pleckstrin's PH domains. SO J-Biol-Chem. 1995 June 16. 270(24). p 14485. AU Philipson-L-H. Kuznetsov-A. Toth-P-T. Murphy-J-F. Szabo-G. Ma-G-H. Miller-R-J. TI Functional expression of an epitope-tagged G protein-coupled K+ channel (GIRK1) SO J-Biol-Chem. 1995 June 16. 270(24). p 14604. AU Koumenis-C. Nunez-Regueiro-M. Raju-U. Cook-R. Eskin-A. TI Identification of three proteins in the eye of Aplysia, whose synthesis is altered by serotonin (5-HT). Possible involvement of these proteins in the ocular circadian system. SO J-Biol-Chem. 1995 June 16. 270(24). p 14619. AU Kimura-K. White-B-H. Sidhu-A. TI Coupling of human D-1 dopamine receptors to different guanine nucleotide binding proteins. Evidence that D-1 dopamine receptors can couple to both Gs and Go. SO J-Biol-Chem. 1995 June 16. 270(24). p 14672. AU Kramer-R-M. Roberts-E-F. Hyslop-P-A. Utterback-B-G. Hui-K-Y. Jakubowski-J-A. TI Differential activation of cytosolic phospholipase A2 (cPLA2) by thrombin and thrombin receptor agonist peptide in human platelets. Evidence for activation of cPLA2 independent of the mitogen-activated protein kinases ERK1/2. SO J-Biol-Chem. 1995 June 16. 270(24). p 14816. AU Kalman-V-K. Erdman-R-A. Maltese-W-A. Robishaw-J-D. TI Regions outside of the CAAX motif influence the specificity of prenylation of G protein {gamma} subunits SO J-Biol-Chem. 1995 June 16. 270(24). p 14835. AU Kravchenko-V-V. Pan-Z. Han-J. Herbert-J-M. Ulevitch-R-J. Ye-R-D. TI Platelet-activating factor induces NF-{kappa}B activation through a G protein-coupled pathway SO J-Biol-Chem. 1995 June 23. 270(25). p 14928. AU Brown-H-A. Gutowski-S. Kahn-R-A. Sternweis-P-C. TI Partial purification and characterization of Arf-sensitive phospholipase D from porcine brain SO J-Biol-Chem. 1995 June 23. 270(25). p 14935. AU Singer-W-D. Brown-H-A. Bokoch-G-M. Sternweis-P-C. TI Resolved phospholipase D activity is modulated by cytosolic factors other than Arf SO J-Biol-Chem. 1995 June 23. 270(25). p 14944. AU Schwenk-U. Schroder-J-M. TI 5-Oxo-eicosanoids are potent eosinophil chemotactic factors. Functional characterization and structural requirements. SO J-Biol-Chem. 1995 June 23. 270(25). p 15029. AU Rossier-M-F. Aptel-H-B-C. Python-C-P. Burnay-M-M. Vallotton-M-B. Capponi-A-M. TI Inhibition of low threshold calcium channels by angiotensin II in adrenal glomerulosa cells through activation of protein kinase C SO J-Biol-Chem. 1995 June 23. 270(25). p 15137. AU Offermanns-S. Simon-M-I. TI G{alpha}15 and G{alpha}16 couple a wide variety of receptors to phospholipase C SO J-Biol-Chem. 1995 June 23. 270(25). p 15175. AU Marx-U-C. Austermann-S. Bayer-P. Adermann-K. Ejchart-A. Sticht-H. Walter-S. Schmid-F-X. Jaenicke-R. Forssmann-W-G. Rosch-P. TI Structure of human parathyroid hormone 1-37 in solution SO J-Biol-Chem. 1995 June 23. 270(25). p 15194. AU Emoto-N. Yanagisawa-M. TI Endothelin-converting enzyme-2 is a membrane-bound, phosphoramidon-sensitive metalloprotease with acidic pH optimum SO J-Biol-Chem. 1995 June 23. 270(25). p 15262. AU Sato-M. Kataoka-R. Dingus-J. Wilcox-M. Hildebrandt-J-D. Lanier-S-M. TI Factors determining specificity of signal transduction by G-protein-coupled receptors. Regulation of signal transfer from receptor to G-protein. SO J-Biol-Chem. 1995 June 23. 270(25). p 15269. AU Palczewski-K. Ohguro-H. Premont-R-T. Inglese-J. TI Rhodopsin kinase autophosphorylation. Characterization of site-specific mutations. SO J-Biol-Chem. 1995 June 23. 270(25). p 15294. AU Usdin-T-B. Gruber-C. Bonner-T-I. TI Identification and functional expression of a receptor selectively recognizing parathyroid hormone, the PTH2 receptor SO J-Biol-Chem. 1995 June 30. 270(26). p 15455. AU Sanada-K. Kokame-K. Yoshizawa-T. Takao-T. Shimonishi-Y. Fukada-Y. TI Role of heterogeneous N-terminal acylation of recoverin in rhodopsin phosphorylation SO J-Biol-Chem. 1995 June 30. 270(26). p 15459. AU Vogel-W-K. Mosser-V-A. Bulseco-D-A. Schimerlik-M-I. TI Porcine m2 muscarinic acetylcholine receptor-effector coupling in Chinese hamster ovary cells SO J-Biol-Chem. 1995 June 30. 270(26). p 15485. AU Li-S. Okamoto-T. Chun-M. Sargiacomo-M. Casanova-J-E. Hansen-S-H. Nishimoto-I. Lisanti-M-P. TI Evidence for a regulated interaction between heterotrimeric G proteins and caveolin SO J-Biol-Chem. 1995 June 30. 270(26). p 15693. AU Abrahamsen-N. Lundgren-K. Nishimura-E. TI Regulation of glucagon receptor mRNA in cultured primary rat hepatocytes by glucose and cAMP SO J-Biol-Chem. 1995 June 30. 270(26). p 15853. AU Kaufman-D-L. Keith-D-E-Jr.. Anton-B. Tian-J. Magendzo-K. Newman-D. Tran-T-H. Lee-D-S. Wen-C. Xia-Y-R. Lusis-A-J. Evans-C-J. TI Characterization of the murine {mu} opioid receptor gene SO J-Biol-Chem. 1995 June 30. 270(26). p 15877. AU Mende-U. Schmidt-C-J. Yi-F. Spring-D-J. Neer-E-J. TI The G protein {gamma} subunit. Requirements for dimerization with {beta} subunits. SO J-Biol-Chem. 1995 June 30. 270(26). p 15892. AU DeMartino-J-A. Konteatis-Z-D. Siciliano-S-J. Riper-G-V. Underwood-D-J. Fischer-P-A. Springer-M-S. TI Arginine 206 of the C5a receptor is critical for ligand recognition and receptor activation by C-terminal hexapeptide analogs SO J-Biol-Chem. 1995 July 7. 270(27). p 15966. AU Ji-I. Ji-T-H. TI Differential roles of exoloop 1 of the human follicle-stimulating hormone receptor in hormone binding and receptor activation SO J-Biol-Chem. 1995 July 7. 270(27). p 15970. AU Wu-D. Kuang-Y. Wu-Y. Jiang-H. TI Selective coupling of {beta}2-adrenergic receptor to hematopoietic-specific G proteins SO J-Biol-Chem. 1995 July 7. 270(27). p 16008. AU Migeon-J-C. Thomas-S-L. Nathanson-N-M. TI Differential coupling of m2 and m4 muscarinic receptors to inhibition of adenylyl cyclase by Gi{alpha} and Go{alpha} subunits SO J-Biol-Chem. 1995 July 7. 270(27). p 16070. AU Hayslett-J-P. Macala-L-J. Smallwood-J-I. Kalghatgi-L. Gassala-Herraiz-J. Isales-C. TI Vasopressin-stimulated electrogenic sodium transport in A6 cells is linked to a Ca2+-mobilizing signal mechanism SO J-Biol-Chem. 1995 July 7. 270(27). p 16082. AU Negishi-M. Irie-A. Sugimoto-Y. Namba-T. Ichikawa-A. TI Selective coupling of prostaglandin E receptor EP3D to Gi and Gs through interaction of {alpha}-carboxylic acid of agonist and arginine residue of seventh transmembrane domain SO J-Biol-Chem. 1995 July 7. 270(27). p 16122. AU Klenchin-V-A. Calvert-P-D. Bownds-M-D. TI Inhibition of rhodopsin kinase by recoverin. Further evidence for a negative feedback system in phototransduction. SO J-Biol-Chem. 1995 July 7. 270(27). p 16147. AU Ishii-K. Gerszten-R. Zheng-Y-W. Welsh-J-B. Turck-C-W. Coughlin-S-R. TI Determinants of thrombin receptor cleavage. Receptor domains involved, specificity, and role of the P3 aspartate. SO J-Biol-Chem. 1995 July 7. 270(27). p 16435. AU Combadiere-C. Ahuja-S-K. Murphy-P-M. TI Cloning and functional expression of a human eosinophil CC chemokine receptor SO J-Biol-Chem. 1995 July 14. 270(28). p 16491. AU Luttrell-L-M. Biesen-T-. Hawes-B-E. Koch-W-J. Touhara-K. Lefkowitz-R-J. TI G{beta}{gamma} subunits mediate mitogen-activated protein kinase activation by the tyrosine kinase insulin-like growth factor 1 receptor SO J-Biol-Chem. 1995 July 14. 270(28). p 16495. AU Monk-P-N. Barker-M-D. Partridge-L-J. Pease-J-E. TI Mutation of glutamate 199 of the human C5a receptor defines a binding site for ligand distinct from the receptor N terminus SO J-Biol-Chem. 1995 July 14. 270(28). p 16625. AU Chueh-S-H. Song-S-L. Liu-T-Y. TI Heterologous desensitization of opioid-stimulated Ca2+ increase by bradykinin or ATP in NG108-15 cells SO J-Biol-Chem. 1995 July 14. 270(28). p 16630. AU Teti-A. Paniccia-R. Goldring-S-R. TI Calcitonin increases cytosolic free calcium concentration via capacitative calcium influx SO J-Biol-Chem. 1995 July 14. 270(28). p 16666. AU Wang-C. Jayadev-S. Escobedo-J-A. TI Identification of a domain in the angiotensin II type 1 receptor determining Gq coupling by the use of receptor chimeras SO J-Biol-Chem. 1995 July 14. 270(28). p 16677. AU Sealfon-S-C. Chi-L. Ebersole-B-J. Rodic-V. Zhang-D. Ballesteros-J-A. Weinstein-H. TI Related contribution of specific helix 2 and 7 residues to conformational activation of the serotonin 5-HT2A receptor SO J-Biol-Chem. 1995 July 14. 270(28). p 16683. AU Smith-J-B. Herschman-H-R. TI Glucocorticoid-attenuated response genes encode intercellular mediators, including a new C-X-C chemokine SO J-Biol-Chem. 1995 July 14. 270(28). p 16756. AU Palmer-T-M. Gettys-T-W. Stiles-G-L. TI Differential interaction with and regulation of multiple G-proteins by the rat A3 adenosine receptor SO J-Biol-Chem. 1995 July 14. 270(28). p 16895. AU Touhara-K. Koch-W-J. Hawes-B-E. Lefkowitz-R-J. TI Mutational analysis of the pleckstrin homology domain of the {beta}-adrenergic receptor kinase. Differential effects on G{beta}{gamma} and phosphatidylinositol 4,5-bisphosphate binding. SO J-Biol-Chem. 1995 July 14. 270(28). p 17000. AU Hawes-B-E. Biesen-T-. Koch-W-J. Luttrell-L-M. Lefkowitz-R-J. TI Distinct pathways of Gi- and Gq-mediated mitogen-activated protein kinase activation SO J-Biol-Chem. 1995 July 21. 270(29). p 17148. AU Wise-A. Lee-T-W. MacEwan-D-J. Milligan-G. TI Degradation of G11{alpha}/Gq{alpha} is accelerated by agonist occupancy of {alpha}1A/D, {alpha}1B, and {alpha}1C adrenergic receptors SO J-Biol-Chem. 1995 July 21. 270(29). p 17196. AU Gao-J-L. Murphy-P-M. TI Cloning and differential tissue-specific expression of three mouse {beta} chemokine receptor-like genes, including the gene for a functional macrophage inflammatory protein-1{alpha} receptor SO J-Biol-Chem. 1995 July 21. 270(29). p 17494. AU Blin-N. Yun-J. Wess-J. TI Mapping of single amino acid residues required for selective activation of Gq/11 by the m3 muscarinic acetylcholine receptor SO J-Biol-Chem. 1995 July 28. 270(30). p 17741. AU Holz-G-G-IV. Leech-C-A. Habener-J-F. TI Activation of a cAMP-regulated Ca2+-signaling pathway in pancreatic {beta}-cells by the insulinotropic hormone glucagon-like peptide-1 SO J-Biol-Chem. 1995 July 28. 270(30). p 17749. AU Peterson-G-L. Toumadje-A. Johnson-W-C-Jr.. Schimerlik-M-I. TI Purification of recombinant porcine m2 muscarinic acetylcholine receptor from Chinese hamster ovary cells. Circular dichroism spectra and ligand binding properties. SO J-Biol-Chem. 1995 July 28. 270(30). p 17808. AU Tseng-M-J. Coon-S. Stuenkel-E. Struk-V. Logsdon-C-D. TI Influence of second and third cytoplasmic loops on binding, internalization, and coupling of chimeric bombesin/m3 muscarinic receptors SO J-Biol-Chem. 1995 July 28. 270(30). p 17884. AU Busch-S. Wieland-T. Esche-H. Jakobs-K-H. Siffert-W. TI G protein regulation of the Na+/H+ antiporter in Xenopus laevis oocytes. Involvement of protein kinases A and C. SO J-Biol-Chem. 1995 July 28. 270(30). p 17898. AU Freedman-N-J. Liggett-S-B. Drachman-D-E. Pei-G. Caron-M-G. Lefkowitz-R-J. TI Phosphorylation and desensitization of the human {beta}1-adrenergic receptor. Involvement of G protein-coupled receptor kinases and cAMP-dependent protein kinase. SO J-Biol-Chem. 1995 July 28. 270(30). p 17953. AU Schoneberg-T. Liu-J. Wess-J. TI Plasma membrane localization and functional rescue of truncated forms of a G protein-coupled receptor SO J-Biol-Chem. 1995 July 28. 270(30). p 18000. AU Chen-C-K. Inglese-J. Lefkowitz-R-J. Hurley-J-B. TI Ca2+-dependent interaction of recoverin with rhodopsin kinase SO J-Biol-Chem. 1995 July 28. 270(30). p 18060. AU Kolakowski-L-F-Jr.. Lu-B. Gerard-C. Gerard-N-P. TI Probing the message:address sites for chemoattractant binding to the C5a receptor. Mutagenesis of hydrophilic and proline residues within the transmembrane segments. SO J-Biol-Chem. 1995 July 28. 270(30). p 18077. AU Cusack-B. McCormick-D-J. Pang-Y-P. Souder-T. Garcia-R. Fauq-A. Richelson-E. TI Pharmacological and biochemical profiles of unique neurotensin 8-13 analogs exhibiting species selectivity, stereoselectivity, and superagonism SO J-Biol-Chem. 1995 August 4. 270(31). p 18359. AU Chen-Y-G. Danoff-A. Shields-D. TI The propeptide of anglerfish preprosomatostatin-I rescues prosomatostatin-II from intracellular degradation SO J-Biol-Chem. 1995 August 4. 270(31). p 18598. AU Prasad-M-V-V-S-V. Dermott-J-M. Heasley-L-E. Johnson-G-L. Dhanasekaran-N. TI Activation of Jun kinase/stress-activated protein kinase by GTPase-deficient mutants of G{alpha}12 and G{alpha}13 SO J-Biol-Chem. 1995 August 4. 270(31). p 18655. AU Chuang-T-T. LeVine-H-III. Blasi-A-D. TI Phosphorylation and activation of {beta}-adrenergic receptor kinase by protein kinase C SO J-Biol-Chem. 1995 August 4. 270(31). p 18660. AU Butkerait-P. Zheng-Y. Hallak-H. Graham-T-E. Miller-H-A. Burris-K-D. Molinoff-P-B. Manning-D-R. TI Expression of the human 5-hydroxytryptamine1A receptor in Sf9 cells. Reconstitution of a coupled phenotype by co-expression of mammalian G protein subunits. SO J-Biol-Chem. 1995 August 4. 270(31). p 18691. -- Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Wed Jul 19 23:00:00 1995 Path: biosci!bms.com!watson_j From: watson_j@bms.com (John Watson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Desperately Seeking Roommate For Neuroscience Date: 20 Jul 1995 07:31:47 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 22 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net If you are going to the Neuroscience Annual meeting in San Diego Nov. 11-16 and would like a roommate to help keep hotel costs down, please email me ASAP -- hotels are filling up there! AJW -------------------------------------------------------------------------- A. John Watson, Ph.D. __ __ __ Bristol-Myers Squibb Pharmaceutical Reseach Institute /__/__/__/\ Central Nervous System Biology, Department 404 /__/__/__/\/\ 5 Research Parkway /__/__/__/\/\/\ Wallingford, CT 06492 \__\__\__\/\/\/ \__\__\__\/\/ (203) 284-6745 \__\__\__\/ (203) 284-7569 fax watson_j@bms.com -------------------------------------------------------------------------- From owner-7tms_r@net.bio.net Wed Jul 19 23:00:00 1995 Path: biosci!rutgers!goliath.montclair.edu!newsserver.jvnc.net!synapse.bms.com!NewsWatcher!user From: watson_j@bms.com (A. John Watson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Rommate for Neuroscience Annual meeting? Date: Thu, 20 Jul 1995 11:38:08 -0300 Organization: Bristol-Myers Squibb Lines: 23 Message-ID: NNTP-Posting-Host: 140.176.107.223 I sent this to the mailing list, but don't know if it gets posted here. Sorry for the bandwidth waste if it gets duplicated. If you are going to the Neuroscience Annual meeting in San Diego Nov. 11-16 and would like a roommate to help keep hotel costs down (or if you know anybody who is), please email me ASAP -- hotels are filling up there! AJW -- __ __ __ /__/__/__/\ A. John Watson, Ph.D. /__/__/__/\/\ Bristol-Myers Squibb Company /__/__/__/\/\/\ CNS Biology, Dept. 404 \__\__\__\/\/\/ Wallingford, CT 06492 \__\__\__\/\/ watson_j@bms.com \__\__\__\/ My opinions, of course, are my own. Which is just as well since nobody else seems to want them. | From owner-7tms_r@net.bio.net Wed Jul 19 23:00:00 1995 Path: biosci!OSU.EDU!beall.3 From: beall.3@OSU.EDU (Clifford Beall) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Yeast expression? Date: 20 Jul 1995 10:49:01 -0700 Organization: Ohio State University Lines: 24 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507201747.NAA23086@bottom.magnus.acs.ohio-state.edu> NNTP-Posting-Host: net.bio.net Dear 7TMS workers, I would like to know other people's experiences with the use of yeast expression systems for 7TMS receptors. Has anyone else gotten good expression by western blots but been unable to demonstrate specific binding of ligand to membranes? We have basically followed the procedures given in the following papers: Marjamaki A et al, "Similar ligand binding in recombinant human alpha 2 C2-adrenoceptors produced in mammalian, insect and yeast cells." European Journal of Pharmacology 1994 Mar 15;267(1):117-21 Sander P et al, "Constitutive expression of the human D2S-dopamine receptor in the unicellular yeast Saccharomyces cerevisiae." Biochimica et Biophysica Acta 1994 Aug 3;1193(2):255-62 Does anyone have variations or tricks to share? Sincerely yours, Cliff Beall Neurobiotechnology Center The Ohio State University Beall.3@osu.edu From owner-7tms_r@net.bio.net Thu Jul 20 23:00:00 1995 Path: biosci!daresbury!nntp-trd.UNINETT.no!trane.uninett.no!Norway.EU.net!EU.net!howland.reston.ans.net!usc!comserv-f-36.usc.edu!user From: bfry@hsc.usc.edu (Bryan Fry) Newsgroups: bionet.molbio.proteins.7tms_r Subject: APOPTOSIS Date: Fri, 21 Jul 1995 16:33:15 -0800 Organization: USC Cancer Center Lines: 8 Sender: bfry@comserv-f-36.usc.edu Message-ID: NNTP-Posting-Host: comserv-f-36.usc.edu This is Bryan Fry at USC Cancer Center, I am writing to inquire if anyone out there has any brilliant ideas on how to induce apoptosis in human breast cancer cells (cell line:MDA-MB-435). We need to induce it in them to use as a control that we are doing involving disintegrins. With regards, Bryan From owner-7tms_r@net.bio.net Wed Jul 26 23:00:00 1995 Path: biosci!rutgers!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 for Aug 11 JBC Date: 28 Jul 1995 01:05:02 GMT Organization: Mass. General Hospital, Renal Unit Lines: 56 Distribution: world Message-ID: NNTP-Posting-Host: receptor.mgh.harvard.edu AU Carman-G-M. Deems-R-A. Dennis-E-A. TI Lipid signaling enzymes and surface dilution kinetics SO J-Biol-Chem. 1995 August 11. 270(32). p 18711. AU Sugaya-T. Nishimatsu-S-. Tanimoto-K. Takimoto-E. Yamagishi-T. Imamura-K. Goto-S. Imaizumi-K. Hisada-Y. Otsuka-A. Uchida-H. Sugiura-M. Fukuta-K. Fukamizu-A. Murakami-K. TI Angiotensin II type 1a receptor-deficient mice with hypotension and hyperreninemia SO J-Biol-Chem. 1995 August 11. 270(32). p 18719. AU Sakai-H. Kumano-E. Ikari-A. Takeguchi-N. TI A gastric housekeeping Cl- channel activated via prostaglandin EP3 receptor-mediated Ca2+/nitric oxide/cGMP pathway SO J-Biol-Chem. 1995 August 11. 270(32). p 18781. AU Knol-J-C. Slik-A-R-. Kesteren-E-R-. Planta-R-J. Heerikhuizen-H-. Vreugdenhil-E. TI A novel G protein {alpha} subunit containing atypical guanine nucleotide-binding domains is differentially expressed in a molluscan nervous system SO J-Biol-Chem. 1995 August 11. 270(32). p 18804. AU Zhou-W. Rodic-V. Kitanovic-S. Flanagan-C-A. Chi-L. Weinstein-H. Maayani-S. Millar-R-P. Sealfon-S-C. TI A locus of the gonadotropin-releasing hormone receptor that differentiates agonist and antagonist binding sites SO J-Biol-Chem. 1995 August 11. 270(32). p 18853. AU Tseng-M-J. Detjen-K. Struk-V. Logsdon-C-D. TI Carboxyl-terminal domains determine internalization and recycling characteristics of bombesin receptor chimeras SO J-Biol-Chem. 1995 August 11. 270(32). p 18858. AU Boie-Y. Sawyer-N. Slipetz-D-M. Metters-K-M. Abramovitz-M. TI Molecular cloning and characterization of the human prostanoid DP receptor SO J-Biol-Chem. 1995 August 11. 270(32). p 18910. AU Bhat-G-J. Thekkumkara-T-J. Thomas-W-G. Conrad-K-M. Baker-K-M. TI Activation of the STAT pathway by angiotensin II in T3CHO/AT1A cells. Cross-talk between angiotensin II and interleukin-6 nuclear signaling. SO J-Biol-Chem. 1995 August 11. 270(32). p 19059. AU Giannini-E. Brouchon-L. Boulay-F. TI Identification of the major phosphorylation sites in human C5a anaphylatoxin receptor in vivo SO J-Biol-Chem. 1995 August 11. 270(32). p 19166. -- Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Wed Jul 26 23:00:00 1995 Path: biosci!RISC.IDG.FI.CNR.IT!giolitti From: giolitti@RISC.IDG.FI.CNR.IT (Alessandro Giolitti) Newsgroups: bionet.molbio.proteins.7tms_r Subject: B1 receptor Date: 27 Jul 1995 23:10:18 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I am again on the bradykinin B1 receptor. Even if the low (36%) homology with the B2 is not an issue, what makes me wonder about the definition of B1 is the fact that actually seems not to be a bradykinin receptor: bk has a very low affinity for it, and its natural ligand is not bk. So should not it to be classified (and dealth with) in a different way? Thanks Alessandro Giolitti _______________________________________ Dr.Alessandro Giolitti Drug Design Dept. Menarini s.r.l. Via Sette Santi, 3 50131 Firenze Italy Ph. : +39-55-5680240 FAX : +39-55-5680419 e-mail: giolitti@risc.idg.fi.cnr.it _______________________________________ From owner-7tms_r@net.bio.net Fri Jul 28 23:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: intracellular 7TM receptors? Date: 29 Jul 1995 19:40:04 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 69 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507300238.WAA23265@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > Paul.Ernsberger@slc5.INS.CWRU.Edu, "Ph.D. (pre@PO.CWRU.EDU) wrote: > > Setting aside the question of mitochondrial receptors, many garden-variety > > 7tms_r are predominantly located intracellularly. This was shown at > > Cornell a few years back by Virginia Pickel and Chiye Aoki (now at NYU) > > who did electron microscopy of beta1 and beta2 adrenergic receptors and > > found a primarily intracellular localization. Many were associated with > > the golgi and were presumably undergoin transport and posttranslational > > modification, but others were in vesicular structures (internalized?). > > If there exists a paper about these observations, could you please post > the reference? I am aware of several electron microscopy studies on the > beta2-AR from the labs of Donny Strosberg and Kai Simons, but I have > never heard of the persons that you have mentioned -- therefore I would > be very interested in any pictures :-) > > /* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ > /* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ > Sorry it took so long to post this list. I had trouble getting it into the right format. Bibliography 1). Aoki C, Zemcik BA, Strader CD, Pickel VM: Cytoplasmic loop of á-adrenergic receptors: Synaptic and intracellular localization and relation to catecholaminergic neurons in the nuclei of the solitary tracts. Brain Res 493:331-347, 1989 2). Aoki C, Pickel VM: Ultrastructural immunocytochemical evidence for presynaptic localization of beta-adrenergic receptors in the striatum and cerebral cortex of rat brain. Ann N Y Acad Sci 604:582-585, 1990 3). Aoki C: á-Adrenergic receptors: Astrocytic localization in the adult visual cortex and their relation to catecholamine axon terminals as revealed by electron microscopic immunocytochemistry. J.Neurosci. 12:781-792, 1992 4). Aoki C, Pickel VM: C-Terminal tail of á-adrenergic receptors: Immunocytochemical localization within astrocytes and their relation to catecholaminergic neurons in N. tractus solitarii and area postrema. Brain Res 571:35-49, 1992 5). Aoki C, Pickel VM: Ultrastructural relations between á-adrenergic receptors and catecholaminergic neurons. Brain Res Bull 29:257-263, 1992 6). Ransn„s LA, Jasper JR, Leiber D, Insel PA: á-Adrenergic-receptor -mediated dissociation and membrane release of the Gs protein in S49 lymphoma-cell membranes. Dependence on Mg2+ and GTP. Biochem J 283:519-524, 1992 7). Roth DA, Urasawa K, Leiber D, Insel PA, Hammond HK: A substantial proportion of cardiac Gs is not associated with the plasma membrane. FEBS Lett 296:46-50, 1992 8). Saffitz JE, Liggett SB: Subcellular distribution of á2-adrenergic receptors delineated with quantitative ultrastructural autoradiography of radioligand binding sites. Circ Res 70:1320-1325, 1992 9). Aoki C, Go C-G, Venkatesan C, Kurose H: Perikaryal and synaptic localization of à2A-adrenergic receptor-like immunoreactivity. Brain Res 650:181-204, 1994 -- 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 Jul 30 23:00:00 1995 Path: biosci!uni-duesseldorf.de!schuessl From: schuessl@uni-duesseldorf.de (Peter Schuessler) Newsgroups: bionet.molbio.proteins.7tms_r Subject: ras on Western blots? Date: 31 Jul 1995 09:58:07 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Distribution: world Message-ID: <199507311657.JAA23277@net.bio.net> NNTP-Posting-Host: net.bio.net Hallo, We work on signal transduction in endoparasites. On Western blots, I could detect ras with an antiserum from human. Interestingley, on these blots in addition to the expected band of about 21kD, a second band appears in the size range of 45kD. After separation in soluble and membrane protein, the 21kD band appears only in the soluble fraction whereas the 45kD band reacts only with the membrane fraction. Is there anybody out there who could explain these observations? Does ras form dimers or exist a protein of about 45kD that is simillar to ras? Could anybody give me a good hint? Thank you for your friendly help Peter Schuessler Peter Schuessler Institut fuer Genetik Heinrich-Heine-Universitaet Duesseldorf D-40225 Duesseldorf Germany Tel: ++49-(0)211-311-2408 e-mail: schuessl@mail.rz.uni-duesseldorf.de From owner-7tms_r@net.bio.net Mon Jul 31 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!newsserver.jvnc.net!netnews.sb.com!news From: Steven_Mcclue-1%notes@sb.com (Steve Mcclue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: ras on Western blots? Date: 1 Aug 1995 08:31:23 GMT Organization: Smithkline Beecham Pharmaceuticals Lines: 25 Distribution: world Message-ID: <3vkoor$ce@phunn1.sb.com> References: <199507311657.JAA23277@net.bio.net> NNTP-Posting-Host: had542.uk.sb.com Mime-Version: 1.0 X-Newsreader: WinVN 0.99.2 In article <199507311657.JAA23277@net.bio.net>, schuessl@uni-duesseldorf.de says... > >Hallo, >We work on signal transduction in endoparasites. On Western blots, I could >detect ras with an antiserum from human. Interestingley, on these blots in >addition to the expected band of about 21kD, a second band appears in the >size range of 45kD. After separation in soluble and membrane protein, the >21kD band appears only in the soluble fraction whereas the 45kD band reacts >only with the membrane fraction. >Is there anybody out there who could explain these observations? Does ras >form dimers or exist a protein of about 45kD that is simillar to ras? Could >anybody give me a good hint? >Thank you for your friendly help Peter Schuessler Peter, Those of us who work on signal-transducing G-proteins think of ras as a 'low-molecular-weight G-protein' because it has a MW of about 21kD. The G-proteins we work on are membrane-bound, and generally have MWs of 39-45 kD. In particular, the alpha subunit of Gs has a MW of about 45kD. I don't know what epitope your antibody was raised against, but if it was anywhere in the GTP-binding site, then your Ab could well cross-react with 'high MW-G-proteins' as the GTP-binding domain is highly conserved in both low- and high- MW G-proteins. My guess is that you are seeing Gs. Steve McClue From owner-7tms_r@net.bio.net Mon Jul 31 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!howland.reston.ans.net!news.nic.surfnet.nl!rug.nl!biochem06.chem.rug.nl!bominaar From: bominaar@chem.rug.nl (A.A. Bominaar) Subject: Re: ras on Western blots? Message-ID: Lines: 40 Sender: root@rug.nl (Operator) Nntp-Posting-Host: biochem06.chem.rug.nl Organization: Faculteit Wis- en Natuurwetenschappen References: <199507311657.JAA23277@net.bio.net> Date: Tue, 1 Aug 1995 08:33:03 GMT In article <199507311657.JAA23277@net.bio.net> schuessl@uni-duesseldorf.de (Peter Schuessler) writes: >Hallo, >We work on signal transduction in endoparasites. On Western blots, I could >detect ras with an antiserum from human. Interestingley, on these blots in >addition to the expected band of about 21kD, a second band appears in the >size range of 45kD. After separation in soluble and membrane protein, the >21kD band appears only in the soluble fraction whereas the 45kD band reacts >only with the membrane fraction. >Is there anybody out there who could explain these observations? Does ras >form dimers or exist a protein of about 45kD that is simillar to ras? Could >anybody give me a good hint? >Thank you for your friendly help Peter Schuessler Before anybody could be making educated guesses I think you should provide us with a little more detail on the antibody you are using. As far as I know Ras does not form homodimers (if that is what you mean by dimers), nor is there any protein known of 45 kDa that is similar to Ras, that is if you exclude the alpha-subunits of hetero-trimeric G-proteins. But as far as the latter are concerned I do not know (Yet I do not know them all) of any anti-ras antibody giving cross-reactivity with G-protein alpha subunits. On the other hand I wuoldn't be surprised if you are being bothered by a protein that just happens to contain a comparable epitope, but has nothing to do with Ras or signal-transduction. Another thing that surprises me is your observation that the 21 kDa protein is only found in the soluble fraction. I would expect at least some in the membranes, due to myristylation. best Ton ______________________________________________________________________________ Anthony A. Bominaar Life stinks Dept. of Biochemistry But so does french cheese University of Groningen Yet, that never stopped me from The Netherlands enjoying it. bominaar@chem.rug.nl ------------------------------------------------------------------------------