From owner-7tms_r@net.bio.net Wed Jan 03 22:00:00 1996 Path: biosci!LILLY.COM!SHARP_JOHN_D From: SHARP_JOHN_D@LILLY.COM ("J.D.SHARP 317-276-4268 DC0434") Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 4 Jan 1996 08:08:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HZLZ8LDD5U0001ML@Lilly.com> NNTP-Posting-Host: net.bio.net unsubscribe sharp_john_d@lilly.com 7TMS_r@net.bio.net From: SHARP JOHN D (MCVAX0::FOOT) To: VMS MAIL ADDRESSEE (INT::"7tms_r@net.bio.net") cc: SHARP JOHN D (MCVAX0::FOOT) From owner-7tms_r@net.bio.net Mon Jan 08 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsserver.jvnc.net!netnews.sb.com!news From: Steven_Mcclue-1@sbphrd.com (Steve Mcclue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: adenylyl/adenylate cyclase Date: 9 Jan 1996 13:42:46 GMT Organization: Smithkline Beecham Pharmaceuticals Lines: 7 Message-ID: <4ctrcm$d9v@phunn1.sb.com> NNTP-Posting-Host: had542.ha.uk.sbphrd.com Mime-Version: 1.0 X-Newsreader: WinVN 0.99.2 Can anyone remind me why we should now call this enzyme Adenylyl cyclase rather than Adenylate? Steve PS(I know this is a 12TM, but I was sure someone here would know the answer, anyway) From owner-7tms_r@net.bio.net Mon Jan 08 22:00:00 1996 Path: biosci!novo.dk!byw From: byw@novo.dk (Robert Bywater) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 9 Jan 1996 23:15:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 61 Sender: daemon@net.bio.net Distribution: world Message-ID: <199601100712.IAA16092@bach> NNTP-Posting-Host: net.bio.net Steve Mcclue writes : >Can anyone remind me why we should now call this >enzyme Adenylyl cyclase rather than Adenylate? --- 'Adenylyl' has caught on in the biochemical literature because some enzymes are regulated by attachment of a moiety of adenylate to a tyrosine hydroxyl group through the phosphate group of the nucleotide. The reaction is thus a phosphorYLation, chemically akin to an acYLation, so the modification of the protein is referred to as an adneylYLation. Now, cyclization of the phosphate group in AMP to cAMP probably involves an 'adenylylation' of a nucleophilic group in the active site of the cyclase, but that is not the point. The net result is that a monoester of phosphoric acid gets converted into a (cyclic) diester. ( The nucleotide itself gets 'adenylylated' ). But I would prefer to say that the object of the verb 'cyclize' is the monoester, which happens to be called adenylic acid, or more properly, as Steve Mcclue writes, adenylate. The adenylate gets cyclized. Therefore, Steve Mcclue should be congratulated for calling into question this erroneous piece of nomenclature. This is not a trivial point because there are questions of chemistry, and grammar, at stake. Robert Bywater ****************************** * * * Robert Bywater * * * * Biostructure Department * * NOVO NORDISK A/S * * * NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB The address below will change soon, please watch this space * * * DK-2880 BAGSVAERD Denmark * * * * email byw@novo.dk * NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB NB The fax number below will change soon, please watch this space * * * fax :: +45 4442 1400 * ****************************** From owner-7tms_r@net.bio.net Tue Jan 09 22:00:00 1996 Path: biosci!WH.BAYER.COM!buggy From: buggy@WH.BAYER.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: adenylyl/adenylate cyclase Date: 10 Jan 1996 09:49:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 6 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601101744.AA13715@mrcs1> NNTP-Posting-Host: net.bio.net I'm still spelling it adenylyl. Because I'm a rebel without a cause on a one-way joyride straight to hell. So there. From owner-7tms_r@net.bio.net Tue Jan 09 22:00:00 1996 Path: biosci!MED.UNC.EDU!erweiss From: erweiss@MED.UNC.EDU ("Ellen R. Weiss") Newsgroups: bionet.molbio.proteins.7tms_r Subject: re: adenylyl/adenylate cyclase Date: 10 Jan 1996 13:26:03 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 25 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601102124.AA26900@med.unc.edu> Reply-To: "Ellen R. Weiss" NNTP-Posting-Host: net.bio.net Steven Mcclue asked the following: Can anyone remind me why we should now call this enzyme Adenylyl cyclase rather than Adenylate? ---------------------------------- I can only relate the following experience. I was at a Cold Spring Harbor meeting on G proteins organized by Michael Wigler, I think in 1986. Someone stood up and said we should resolve the conflict between calling guanine nucleotide binding proteins "N proteins" or "G proteins". Several people argued in favor of "G proteins". After that, Lutz Birnbaumer stood up and said he would go for "G proteins" if everyone would agree to "adenlylyl cyclase" rather than "adenylate cyclase". Everyone agreed, and it seemed that all the papers published after that meeting used that nomenclature. Ellen Weiss University of North Carolina From owner-7tms_r@net.bio.net Tue Jan 09 22:00:00 1996 Path: biosci!MC.DUKE.EDU!premo003 From: premo003@MC.DUKE.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: bacteriorhodopsin Date: 10 Jan 1996 07:00:44 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HZUAI5ZX7I002WU8@mc.duke.edu> NNTP-Posting-Host: net.bio.net Dear 7TMers: Someone in our group is looking for a source for bacteriorhodopsin protein. I know I have seen purple membranes for sale in a catalog a couple of years ago, but can't remember who sells them. Does anyone out there have any info? Thanks much! Richard Premont LefkoLand Department of Medicine Duke University Medical Center Durham, North Carolina 27710 (919) 684-2974 FAX -8875 From owner-7tms_r@net.bio.net Tue Jan 09 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!psgrain!nntp.teleport.com!usenet From: nevek@teleport.com (Kim Neve) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: adenylyl/adenylate cyclase Date: Wed, 10 Jan 1996 19:13:03 GMT Organization: Oregon Health Sciences University Lines: 15 Message-ID: <4d0ot4$hl4@maureen.teleport.com> References: <4ctrcm$d9v@phunn1.sb.com> NNTP-Posting-Host: ip-pdx03-30.teleport.com X-Newsreader: Forte Free Agent 1.0.82 Steven_Mcclue-1@sbphrd.com (Steve Mcclue) wrote: >Can anyone remind me why we should now call this >enzyme Adenylyl cyclase rather than Adenylate? I think that the primary reason is because Gilman said that we should. If you check the enzyme nomenclature web site, at http://expasy.hcuge.ch/sprot/enzyme.html, you will find that adenylate cyclase is listed as the standard name. Kim Neve From owner-7tms_r@net.bio.net Wed Jan 10 22:00:00 1996 Path: biosci!bcm.tmc.edu!pendragon!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!newsfeed.internetmci.com!globe.indirect.com!usenet From: coburn@indirect.com (anne coburn) Newsgroups: bionet.molbio.proteins.7tms_r Subject: PROTEIN CHEMIST OPENING Date: 11 Jan 1996 22:17:23 GMT Organization: Coburn Scientific Lines: 33 Sender: -Not-Authenticated-[3112] Message-ID: <4d429j$59h@globe.indirect.com> NNTP-Posting-Host: s34.tucslip.indirect.com X-Posted-From: InterNews 1.0.7@s34.tucslip.indirect.com Xdisclaimer: No attempt was made to authenticate the sender's name. A leading biotechnology company in Southern California seeks a: RESEARCH ASSOCIATE II/III - PROTEIN CHEMISTRY This company is at the forefront in the discovery, expression, purification, characterization and laboratory-scale development of new protein therapeutics. If you have specialized knowledge PROTEIN CHEMISTRY and PROTEIN SEQUENCING, this is an opporutnity for you to enhance your career in an environment where scientists have the resources and support necessary to develop their full potential. This is an immediate opening, com- pensation is competitive and afforable housing is available. REQUIREMENTS --- MS in Chemistry, Biochemistry or related field. --- 5+ years experience in a research environment. --- Expertise in PROTEIN SEQUENCING and MODIFICATION, HPLC and gel electrophoresis, and peptide mapping. --- The ability to operate specialized laboratory equipment and computers. --- Strong analytical skills and a knowledge of good laboratory techniques. --- Proven interpersonal and communication skills. --- The ability to work independently with minimum supervision. For immediate and confidential consideration, contact ANNE COBURN Coburn Search & Consulting 4449 East Whitman Tucson, AZ 85711-3515 Phone: 520-881-0084 Fax: 520-321-1475 e-mail: coburn@indirect.com From owner-7tms_r@net.bio.net Thu Jan 11 22:00:00 1996 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: BIOSCI miniFAQ, ver. 14-DEC-95 Date: 12 Jan 1996 02:00:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 199 Sender: daemon@net.bio.net Distribution: world Message-ID: <199601121000.CAA20143@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 14-DEC-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. Contents: -------- 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index in addition to the master index for the entire set. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Sun Jan 14 22:00:00 1996 Path: biosci!MEDUSA.UNM.EDU!lsklar From: lsklar@MEDUSA.UNM.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Position available Date: 15 Jan 1996 06:14:50 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 51 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Research Associate Position To investigate the relationship of formyl peptide ligand-receptor-G protein dynamics to cell activation and cell physiology. We are interested in the implications of receptor-G protein precoupling for signal transduction. PhD, MD with research experience or related degree and related experience required. Desirable qualifications include experience in signal transduction, fluorescence, and a record of publications. For best consideration, apply by March 1,1996. Position will remain open until filled. Opportunities include: 1) real-time subsecond analysis of ligand-receptor interactions with stopped flow flow cytometry and spectroscopy; 2) analysis of a family of fluorescent ligands with variable kinetic characteristics; 3) analysis of mutant receptors; 4) biochemical analysis of transduction assemblies including phosphorylation; 5) computer modeling; 6) access to the Los Alamos National Flow Cytometry Resource Contact or send CV to: Larry A. Sklar, PhD Professor of Pathology, UNM Health Sciences Center Co-Director National Flow Cytometry Resource, Los Alamos Nat. Lab Phone 505-277-7249; Fax 505-255-7790 Address: Cancer Center Rm 325 900 Camino de Salud University of New Mexico Health Sciences Center Albuquerque, NM 87131 Representative recent publications 1. Neubig, R. and Sklar, L.A., Subsecond modulation of formyl peptide linked guanine nucleotide-binding proteins by guanosine 5'-0-(3-thio) triphosphate in permeabilized neutrophils, Mol. Pharm, 43:734-740, 1993. 2. Posner, R.G., Fay, S.P., Domalewski, M. and Sklar, L.A., Continuous spectrofluorometric analysis of formyl peptide receptor ternary complex interactions. Mol. Pharm., 45:65-73, 1994. 3. Nolan, J.P., Posner, R.G., Martin, J.C., Habbersett, R., and Sklar, L.A. A Rapid Mix Flow Cytometer With Subsecond Kinetic Resolution, Cytometry, 21:223-229,1995. 4. Domalewski, M.D., Guyer, D.A., Freer, R.J., Muthukumaraswamy, N. and Sklar,L.A. Fixation traps receptors in high and low affinity forms that can be regulated by GTP[S] in the absence of ligand, In Press, J. Rec. Res. 1996 From owner-7tms_r@net.bio.net Mon Jan 15 22:00:00 1996 Path: biosci!ITSA.UCSF.EDU!barber From: barber@ITSA.UCSF.EDU (Diane Barber) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 16 Jan 1996 17:40:53 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Does anyone know whether and how COS1 and COS7 cells are different? Diane Barber From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!MOLDEV.COM!jack_owicki From: jack_owicki@MOLDEV.COM (Jack Owicki) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Endogenous GPCRs Date: 17 Jan 1996 17:44:06 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 60 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net <(In reply to Rick Longton's query: Does anyone know of a listing of endogenous GPCRs in HEK 293, CHO and COS cells? > We've done a short survey of functional responses of several cell lines to a variety of GPCR agonists, using our Cytosensor=AE Microphysiometer to observe receptor activation via subsequent perturbations of cellular metabolic activity (see McConnell et al.(1992) Science257:1906; Owicki et al. (1994) Ann. Rev. Biophys. Biomol. Struct. 23:87). These are unpublished data based on a two-day experiment by Greg Baxter, not meant to be an exhaustive survey and not highly replicated (standard disclaimer here!), but I thought you might find them useful nonetheless. Cell lines tested: CHO-K1, L(tk-), HeLa, 3T3, HEK-293. Agonists tested: 5HT (1 =B5M); Octopamine (10 =B5M); Endothelin-1 (100 nM); Bradykinin (10 = =B5M); Angiotensin-II (10 =B5M); SKF-38393 (10 =B5M; dopamine D1 selective); quinpirole (10 =B5M; dopamine D2 selective); Adenosine (10 =B5M); Met-enkephalin (10 =B5M); Isoproterenol (10 =B5M; beta adrenergic selective)= ; Carbachol (100 =B5M; muscarinic and nicotinic); ATP (10 =B5M; purinoceptor). All combinations of cells and agonists were tested. The following showed functional responses ("?" indicates a weak apparent response of questionable significance): CHO-K1: 5HT, ATP L(tk-): Adenosine (?), ATP HeLa: Endothelin (?), Isoproterenol (?), Carbachol, ATP 3T3: Endothelin, Adenosine (?), Isoproterenol, ATP HEK-293: Endothelin (?), Adenosine, Isoproterenol, Carbachol, ATP In separate studies, we have seen responses to thrombin and PAF in CHO-K1 ce= lls. Our results are pretty consistent with Schonbrunn's original list of endogenous receptors in CHO-K1, HEK-293, and L(tk-) cells, just reposted in reply to Longton. We appear to have obtained responses to all the receptors on that list for which we have tested agonists, with one exception. CHO-K1 cells are reported to express very low levels of beta adrenergic receptors, and we didn't see a response; I'm not aware whether a functional response has ever been demonstrated by any means in this case. ...Jack Owicki ------------------------------------------------------------ John C. Owicki, Ph.D. Associate Technical Director Molecular Devices Corporation 1311 Orleans Drive Sunnyvale, CA 94089 E-mail: jack_owicki@moldev.com Phone: (408) 747-3514; Fax: (408) 747-3601 From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!MC.DUKE.EDU!koch0002 From: koch0002@MC.DUKE.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Post-Doc Position Date: 17 Jan 1996 06:09:30 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I040QGH0NQ000NDP@mc.duke.edu> NNTP-Posting-Host: net.bio.net I have a post-doc position available in my laboratory to study G protein-coupled signaling-based gene therapy approaches to vascular disorders including vein graft intimal hyperplasia and arterial restenosis. Animal models here at Duke are already established to study our novel approach to these problems. Experience in vascular smooth muscle a plus. Interested parties can contact me via email or phone. Walter J. Koch, Ph.D. Assistant Professor Department of Surgery Duke University Medical Center Box 2606 Durham, NC 27710 email: KOCH0002@mc.duke.edu 919-684-3007 FAX: 919-681-5262 From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!FARMR1.MED.UTH.TMC.EDU!aschonb From: aschonb@FARMR1.MED.UTH.TMC.EDU (A. Schonbrunn) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Endogenous GPCRs Date: 17 Jan 1996 15:27:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 59 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net This is the list compiled from the responses that I received to my original quiery about endogenous receptors. If anyone knows of any other receptors in these cells and/or disagrees with what I have listed please let me know. Agi Schonbrunn **************************************************************************** ********************** CHO-K1 - beta adrenergic receptors: =89 30 fmol/mg protein - thrombin receptors coupled to PLC - CCK coupled to PLC - alpha2-adrenergic receptors (<40 fmol/mg). - calcitonin receptors - type I VIP receptors - ATP coupled to Ca++ mobilisation (at 10-5 M) HEK-293 - beta adrenergic receptors: =89 7-20 fmol/mg protein - somatostatin receptors coupled to cyclase inhibition - muscarinic receptors coupled to: - stimulation of PLC (JBC 267:31, 1992) and - inhibition of cylcase (JBC 267: 24858, 1992) - a thrombin receptor - UTP (P2u) receptor - PGE EP2 receptor - adenosine A2b receptor - lysophosphatidic acid receptor COS - beta adrenergic receptors - thrombin receptor - UTP (P2u) receptor - PGE EP2 receptor - adenosine A2b receptor - lysophosphatidic acid receptor Ltk- - P2u (UTP) receptor (PLC & Ca) ************************************************************************** Agi Schonbrunn, Ph.D. Dept. of Pharmacology, Univ. of Texas Medical School >Does anyone know of a listing of endogenous GPCRs in HEK 293, CHO and COS >cells? >I remember someone querying the group on this topic a while ago, but I don'= t >know if anything ever came of it. Thanks for any help. Rick Longton SmithKline Beecham Pharmaceuticals 610-270-7418 Rick_D_Longton@sbphrd.com From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!WH.BAYER.COM!jfs From: jfs@WH.BAYER.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: (none) Date: 17 Jan 1996 06:07:26 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601171402.AA15311@mrcs1> NNTP-Posting-Host: net.bio.net Dear Diane- Cos 7 and Cos 1 cells have the same origin and are identical. They are just two different clones. I have compared them for transfection efficiency and they perform the same. PS I used to work for FSG and BKK (ring any bells). See ya around the net- Jean Schaefer Bayer Research Center West Haven, CT jfs@wh.bayer.com From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!UCDAVIS.EDU!cahaskell From: cahaskell@UCDAVIS.EDU (Chris Haskell) Newsgroups: bionet.molbio.proteins.7tms_r Subject: COS 1 & 7 differences Date: 17 Jan 1996 08:40:12 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 39 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net According to ATCC descriptions, the difference is the line I've indicated below. Otherwise, COS-7 has the same description listed below. I'm sure that there are many more differences in the literature... TCC CRL-1650 COS-1 (Kidney, SV40 transformed, African green monkey) Passage Frozen: Unknown. Current medium for propagation: Dulbecco's modified Eagle's medium, 90%; FBS, 10%. Additional Information: COS-1 is a fibroblast-like cell line established from CV-1 simian cells (ATCC CCL-70) which were transformed by an origin-defective mutant of SV40 which codes for wild-type T antigen. This line contains T antigen, retains complete permissiveness for lytic growth of SV40, supports the replication of ts A209 virus at 40C and supports the replication of pure populations of SV40 mutants with deletions in the early region. -> This line contains a single integrated copy of the complete early region of SV40 DNA. This is a suitable host for transfection, especially for vectors requiring expression of SV40 T antigen (ATCC 37192, 37193, 53100). Handle as potentially biohazardous material under at least Biosafety Level 2 containment. Reference: Cell 23: 175-182, 1981. Submitted by: Y. Gluzman, Cold Spring Harbor Laboratory, Cold Spring Harbor, NY. Price Code: J ----------------------------------------------------------------------- Chris Haskell Dept. Biological Chemistry School of Medicine cahaskell@ucdavis.edu University of California, Davis (916) 752-9070 Davis, CA 95616 ----------------------------------------------------------------------- From owner-7tms_r@net.bio.net Tue Jan 16 22:00:00 1996 Path: biosci!sbphrd.com!Rick_D_Longton From: Rick_D_Longton@sbphrd.com (Rick D Longton) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Endogenous GPCRs Date: 17 Jan 1996 14:19:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 11 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601180119.AA5272@pho903.sbphrd.com> NNTP-Posting-Host: net.bio.net Does anyone know of a listing of endogenous GPCRs in HEK 293, CHO and COS cells? I remember someone querying the group on this topic a while ago, but I don't know if anything ever came of it. Thanks for any help. Rick Longton SmithKline Beecham Pharmaceuticals 610-270-7418 Rick_D_Longton@sbphrd.com From owner-7tms_r@net.bio.net Wed Jan 17 22:00:00 1996 Path: biosci!WELCHLINK.WELCH.JHU.EDU!bjmarg From: bjmarg@WELCHLINK.WELCH.JHU.EDU (BARRY J MARGULIES) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Frank Kolakowski Date: 18 Jan 1996 10:16:51 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Sorry for the broadcast to the whole world, but... Frank, if you're out there, I can't find your new e-mail address (since you moved from MGH). Please e-mail me b/c I have a couple questions about the GCR database. Thanks. We now return you to your regularly scheduled program. -Barry (bjmarg@welchlink.welch.jhu.edu) URL: http://www.welch.jhu.edu/homepages/bjmarg/html/homepage.html xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx GEORGIA CONFEDERATE SOLDIERS from a monument at Antietam National Battlefield, just south We sleep here in obedience to law. of The Cornfield. When duty called, we came. And no, I'm not from Georgia, When country called, we died. I just thought it was cool. xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx From owner-7tms_r@net.bio.net Thu Jan 18 22:00:00 1996 Path: biosci!sbphrd.com!Rick_D_Longton From: Rick_D_Longton@sbphrd.com (Rick D Longton) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Endog. GPCRs Results Date: 19 Jan 1996 07:28:39 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 146 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601191828.AA1406@pho903.sbphrd.com> NNTP-Posting-Host: net.bio.net Here are the results of my recent inquiry regarding endogenous GPCRs in HEK 293, COS and CHO cells. Many thanks to those who replied. Rick Longton SmithKline Beecham Pharmaceuticals 610-270-7418 Rick_D_Longton@sbphrd.com ---------------------------------------------------------------------------------------- ---------------------------------------------------------------------------------------- ---------------------------------------------------------------------------------------- This is the list compiled from the responses that I received to my original quiery about endogenous receptors. If anyone knows of any other receptors in these cells and/or disagrees with what I have listed please let me know. Agi Schonbrunn **************************************************************************** ********************** CHO-K1 - beta adrenergic receptors: 30 fmol/mg protein - thrombin receptors coupled to PLC - CCK coupled to PLC - alpha2-adrenergic receptors (<40 fmol/mg). - calcitonin receptors - type I VIP receptors - ATP coupled to Ca++ mobilisation (at 10-5 M) HEK-293 - beta adrenergic receptors: 7-20 fmol/mg protein - somatostatin receptors coupled to cyclase inhibition - muscarinic receptors coupled to: - stimulation of PLC (JBC 267:31, 1992) and - inhibition of cylcase (JBC 267: 24858, 1992) - a thrombin receptor - UTP (P2u) receptor - PGE EP2 receptor - adenosine A2b receptor - lysophosphatidic acid receptor COS - beta adrenergic receptors - thrombin receptor - UTP (P2u) receptor - PGE EP2 receptor - adenosine A2b receptor - lysophosphatidic acid receptor Ltk- - P2u (UTP) receptor (PLC & Ca) ************************************************************************** Agi Schonbrunn, Ph.D. Dept. of Pharmacology, Univ. of Texas Medical School ------------------------------------------------------- ------------------------------------------------------- ------------------------------------------------------- <(In reply to Rick Longton's query: Does anyone know of a listing of endogenous GPCRs in HEK 293, CHO and COS cells? > We've done a short survey of functional responses of several cell lines to a variety of GPCR agonists, using our Cytosensor=AE Microphysiometer to observe receptor activation via subsequent perturbations of cellular metabolic activity (see McConnell et al.(1992) Science257:1906; Owicki et al. (1994) Ann. Rev. Biophys. Biomol. Struct. 23:87). These are unpublished data based on a two-day experiment by Greg Baxter, not meant to be an exhaustive survey and not highly replicated (standard disclaimer here!), but I thought you might find them useful nonetheless. Cell lines tested: CHO-K1, L(tk-), HeLa, 3T3, HEK-293. Agonists tested: 5HT (1 =B5M); Octopamine (10 =B5M); Endothelin-1 (100 nM); Bradykinin (10 = =B5M); Angiotensin-II (10 =B5M); SKF-38393 (10 =B5M; dopamine D1 selective); quinpirole (10 =B5M; dopamine D2 selective); Adenosine (10 =B5M); Met-enkephalin (10 =B5M); Isoproterenol (10 =B5M; beta adrenergic selective)= ; Carbachol (100 =B5M; muscarinic and nicotinic); ATP (10 =B5M; purinoceptor). All combinations of cells and agonists were tested. The following showed functional responses ("?" indicates a weak apparent response of questionable significance): CHO-K1: 5HT, ATP L(tk-): Adenosine (?), ATP HeLa: Endothelin (?), Isoproterenol (?), Carbachol, ATP 3T3: Endothelin, Adenosine (?), Isoproterenol, ATP HEK-293: Endothelin (?), Adenosine, Isoproterenol, Carbachol, ATP In separate studies, we have seen responses to thrombin and PAF in CHO-K1 ce= lls. Our results are pretty consistent with Schonbrunn's original list of endogenous receptors in CHO-K1, HEK-293, and L(tk-) cells, just reposted in reply to Longton. We appear to have obtained responses to all the receptors on that list for which we have tested agonists, with one exception. CHO-K1 cells are reported to express very low levels of beta adrenergic receptors, and we didn't see a response; I'm not aware whether a functional response has ever been demonstrated by any means in this case. ...Jack Owicki ------------------------------------------------------------ John C. Owicki, Ph.D. Associate Technical Director Molecular Devices Corporation 1311 Orleans Drive Sunnyvale, CA 94089 E-mail: jack_owicki@moldev.com Phone: (408) 747-3514; Fax: (408) 747-3601 ----------------------------------------------- ----------------------------------------------- ----------------------------------------------- Reply to: RE>Endogenous GPCRs Rick, could you post the results of your inquiry. HEK have muscarinic receptors. Stuart Sealfon -------------------------------------- Date: 1/17/96 6:18 PM To: Stuart Sealfon From: Rick D Longton Does anyone know of a listing of endogenous GPCRs in HEK 293, CHO and COS cells? I remember someone querying the group on this topic a while ago, but I don't know if anything ever came of it. Thanks for any help. Rick Longton SmithKline Beecham Pharmaceuticals 610-270-7418 Rick_D_Longton@sbphrd.com From owner-7tms_r@net.bio.net Sun Jan 21 22:00:00 1996 Path: biosci!musc.edu!laniersm From: laniersm@musc.edu (Stephen M Lanier) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 22 Jan 1996 14:27:15 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net A student asked me the following: What types of adenylylcyclase are expressed in L cells (tk-)? Thanks. From owner-7tms_r@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: ritaber@aol.com (RITABER) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Electrophysiologists,Molecular Biologist,Protein Biochemists Jobs Date: 23 Jan 1996 20:12:22 -0500 Organization: America Online, Inc. (1-800-827-6364) Lines: 26 Sender: root@newsbf02.news.aol.com Message-ID: <4e411m$4ua@newsbf02.news.aol.com> Reply-To: ritaber@aol.com (RITABER) NNTP-Posting-Host: newsbf02.mail.aol.com Synaptic Pharmaceutical Corp. has immediate openings for staff positions in our Molecular and Cell Biology Department. Successful candidates will join a multi-disciplinary team focused on the discovery of novel G protein-coupled and kinase receptors expressed in the nervous system. Electrophysiologists Applicants should have demonstrated expertise in Xenopus oocyte electrophysiology and either a PhD degree with 2 years of postdoc experience or an MS degree.. Experience in expression cloning and/or second messenger systems would be beneficial. Molecular Biologists Applicants should have an MS/BS degree with experience in molecular biology. Expression cloning experience is desirable. Protein Biochemist Applicants should have a PhD degree with 2 years of postdoc experience in protein purification and characterization. Experience with protein kinase receptors would be desirable. Synaptic Pharmaceutical offers excellent compensation and benefits that include a 401(k) plan and equity participation programs. For consideration, indicate the position to which you are responding and send or fax your resume in confidence to: Human Resource Manager, Department 170A , Synaptic Pharmaceutical Corp., 215 College Road, Paramus, N. J. 07652. Fax # 201 261 0623. From owner-7tms_r@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!UCONNVM.UCONN.EDU!GMS95003 From: GMS95003@UCONNVM.UCONN.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 23 Jan 1996 10:04:12 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: <960123.130157.EST.GMS95003@UConnVM.UConn.Edu> NNTP-Posting-Host: net.bio.net Hi, I am interested in knowing about the methods of obtaining the transmembrane proteins by cell culture. I know of a method utilising insect cell line cultures, I would like to know about other alternatives. From owner-7tms_r@net.bio.net Mon Jan 22 22:00:00 1996 Path: biosci!UH.EDU!RABond From: RABond@UH.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Inhibitory B-adrs? Date: 23 Jan 1996 08:29:58 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0CHGH7CLE0006K6@Post-Office.UH.EDU> NNTP-Posting-Host: net.bio.net Does anyone know of any reports where the net response to B-adrenoceptor activation is inhibitory at the second messenger? I am aware of the Lakatta data for the B2, but the net effect of receptor activation is still stimulatory. Thanks. Richard A. Bond, Ph.D.: Department of Pharmacological and Pharmaceutical Sciences, University of Houston, 4800 Calhoun, Houston, Texas 77204-5515. E-mail: RABond@UH.edu telephone: (713) 743-1210, fax: (713) 743-1229. From owner-7tms_r@net.bio.net Tue Jan 23 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!zib-berlin.de!fu-berlin.de!news.dfn.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: (none) Date: 23 Jan 1996 22:56:15 GMT Organization: University of Wuerzburg, Germany Lines: 29 Distribution: world Message-ID: <4e3p2f$fsv@winx03.informatik.uni-wuerzburg.de> References: <960123.130157.EST.GMS95003@UConnVM.UConn.Edu> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] GMS95003@UCONNVM.UCONN.EDU wrote: > I am interested in knowing about the methods of obtaining the transmembrane > proteins by cell culture. I know of a method utilising insect cell line > cultures, I would like to know about other alternatives. There have been numerous attempts to produce recombinant seven transmembrane helix receptors. A very recent review is: @article{grisshammer:95, author = {R. Grisshammer and C. G. Tate}, title = {Overexpression of integral membrane proteins for structural studies.}, journal = {Q. Rev. Biophys.}, volume = 28, number = 3, pages = {315--422}, year = 1995 } Recently a group has reported high expression levels of a neurokinin-2 receptor in Pichia pastoris, similar to those regularly obtained in Sf9 (insect) cells. --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP3 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Wed Jan 24 22:00:00 1996 Path: biosci!HLSUN.REDCROSS.ORG!hlatim From: hlatim@HLSUN.REDCROSS.ORG ("Timothy Hla") Newsgroups: bionet.molbio.proteins.7tms_r Subject: Symposium on Bioactive Lipids/ prostanoids/ sphingolipids Date: 25 Jan 1996 10:29:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: <9601251823.AA01322@hlsun.red-cross.org> Reply-To: "Timothy Hla" NNTP-Posting-Host: net.bio.net Frontiers on Bioactive Lipids'96 XVIth Washington International Spring Symposium Washington, DC MAY 6 to 9, 1996 Topics: Eicosanoids, cyclooxygenases, eicosanoid receptors, receptors and signalling by phospholipids, sphingolipids, fatty acids and esters, lipid receptors and antagonists, lipid binding proteins. Speakers: W.L.Smith, P. Needleman, C. DiRusso, PA Grimaldi, WEM Lands, CC Felder, JT O'Flaherty, CN Serhan, JH Exton, TK Harden, WH Molenaar, F Snyder, YA Hannun, RE Pagano, S Spiegel, VA BAnkaitis, NM Bass, J Knudsen, J Storch, JH Veerkamp, AW Ford-Hutchinson, CD Funk, EJ Goetzl, PV Halushka. Please contact: JY Vanderhoek Dept of Biochemistry and Molecular Biology George Washington University 2300 Eye St., NW Washington, DC 20037 ph: 202-994-2929 or -3709 fx: 202-994-8974 e-mail: jyvdh@gwis2.circ.gwu.edu From owner-7tms_r@net.bio.net Wed Jan 24 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer2.itd.umich.edu!newsxfer.itd.umich.edu!news.mtu.edu!msunews!netnews.upenn.edu!brass2.med.upenn.edu!user From: obrien@pharm.med.upenn.edu (PJO`B) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Pichia expression of 7TMS Receptors Followup-To: bionet.molbio.proteins.7tms_r Date: Thu, 25 Jan 1996 13:25:48 -0500 Organization: University of Pennsylvania Lines: 43 Distribution: world Message-ID: References: <960123.130157.EST.GMS95003@UConnVM.UConn.Edu> <4e3p2f$fsv@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: brass2.med.upenn.edu In article <4e3p2f$fsv@winx03.informatik.uni-wuerzburg.de>, krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) wrote: > GMS95003@UCONNVM.UCONN.EDU wrote: > > I am interested in knowing about the methods of obtaining the transmembrane > > proteins by cell culture. I know of a method utilising insect cell line > > cultures, I would like to know about other alternatives. > > There have been numerous attempts to produce recombinant seven > transmembrane helix receptors. A very recent review is: > > @article{grisshammer:95, > author = {R. Grisshammer and C. G. Tate}, > title = {Overexpression of integral membrane proteins for > structural studies.}, > journal = {Q. Rev. Biophys.}, > volume = 28, > number = 3, > pages = {315--422}, > year = 1995 > } > > Recently a group has reported high expression levels of a neurokinin-2 > receptor in Pichia pastoris, similar to those regularly obtained in > Sf9 (insect) cells. > > --Cornelius. Also see: FEBS Letters 377:451-56 (1995) Expression of functional mouse 5-HT5a serotonin receptor in...Pichia pastoris. in this article, "functional" relates to agonist {[H3]-LSD} binding. Immunodetection of myc tagged receptors showed staining in the ER and in cytoplasmic membrane vesicles, but none on the plasma membrane. Hope this helps Peter From owner-7tms_r@net.bio.net Sun Jan 28 22:00:00 1996 Path: biosci!WELCHLINK.WELCH.JHU.EDU!bjmarg From: bjmarg@WELCHLINK.WELCH.JHU.EDU (BARRY J MARGULIES) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Expression in bacteria Date: 29 Jan 1996 16:23:47 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hi, again, folks. I've been told that bacterial expression is a notoriously poor recombinant system for mammalian GCRs (i.e., bacteria have a difficult time making these proteins). My p.i. would like a reference to this, besides "word of mouth," for an upcoming publication. Any help? -Barry (bjmarg@welchlink.welch.jhu.edu) URL: http://www.welch.jhu.edu/homepages/bjmarg/html/homepage.html xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx GEORGIA CONFEDERATE SOLDIERS from a monument at Antietam National Battlefield, just south We sleep here in obedience to law. of The Cornfield. When duty called, we came. And no, I'm not from Georgia, When country called, we died. I just thought it was cool. xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx From owner-7tms_r@net.bio.net Mon Jan 29 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!sgigate.sgi.com!sdd.hp.com!col.hp.com!csn!magnus.acs.ohio-state.edu!lerc.nasa.gov!purdue!oitnews.harvard.edu!das-news2.harvard.edu!fas-news.harvard.edu!green!rice From: rice@green.harvard.edu (Ken Rice) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Lysine in 2nd position of DRY-like motif? Date: 30 Jan 1996 20:13:11 GMT Organization: Organismal and Evolutionary Biology Lines: 10 Message-ID: <4elu4n$oka@decaxp.harvard.edu> NNTP-Posting-Host: green.harvard.edu The DRY motif in the rhodopsin-beta2 family is seen in several variations: DRC, DRF, DRH, ERC, etc. Is anyone aware of a member of the family that has a lysine in the second position of the motif, e.g. DKY, EKC, or whatever? Thanks, Ken Rice From owner-7tms_r@net.bio.net Tue Jan 30 22:00:00 1996 Path: biosci!OCELOT.RUTGERS.EDU!WCUI From: WCUI@OCELOT.RUTGERS.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: 2D and 3D structure of GPCR ligands Date: 31 Jan 1996 09:37:02 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <01I0NQZLRJPKA0URL2@mbcl.rutgers.edu> NNTP-Posting-Host: net.bio.net Hi fellow 7tmer: I like to find sources for 2D and 3D structures of GPCR ligands. Is there already a database that contains such data? if so, where are they and how can I get them? if not, where should I start if I want build such database? Any suggestions will be fully appreciated. Weili Cui, Ph.D. Senior Scientist Synaptic Pharmaceutical Corp. From owner-7tms_r@net.bio.net Tue Jan 30 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!howland.reston.ans.net!plug.news.pipex.net!pipex!tube.news.pipex.net!pipex!dish.news.pipex.net!pipex!news00.sunet.se!sunic!news99.sunet.se!news.funet.fi!news.abo.fi!usenet From: Tuomo Glumoff Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Expression in bacteria Date: 31 Jan 1996 15:30:05 GMT Organization: Turku Centre for Biotechnology Lines: 7 Message-ID: <4eo1tt$s1q@josie.abo.fi> References: NNTP-Posting-Host: tuomo.btk.utu.fi Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.12(Macintosh; I; 68K) X-URL: news:Pine.SOL.3.91.960129191750.25223A-100000@welchlink.welch.jhu.edu You could look at: Grisshammer & Tate, Quarterly Reviews of Biophysics vol 28, pp. 315-422 (1995): "Overexpression of integral membrane proteins for structural studies". Tuomo From owner-7tms_r@net.bio.net Tue Jan 30 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!news.sprintlink.net!news.cc.sunysb.edu!gdpmac4.chem.sunysb.edu!user From: rocket@chem.sunysb.com Newsgroups: bionet.molbio.proteins.7tms_r Subject: RE:Expression in Bacteria Date: 1 Feb 1996 06:01:37 GMT Organization: State University of New York at Stony Brook Lines: 11 Message-ID: NNTP-Posting-Host: gdpmac4.chem.sunysb.edu Here are three references for expression of 7tms receptors in ecoli. Appears you need a secretion leader, but not always... I'm tempted to try, but i don't know... 1. Chapot et al. (1990) Eur. J. Biochem. 187;137-144 2. Freissmuth et al. (1991) PNAS 88;8548-8552. 3. Marullo et al. (1988) PNAS 85;7551-7555. jte johne@gdpiris.chem.sunysb.edu From owner-7tms_r@net.bio.net Wed Jan 31 22:00:00 1996 Path: biosci!CCS.BBK.AC.UK!UBCG91C From: UBCG91C@CCS.BBK.AC.UK Newsgroups: bionet.molbio.proteins.7tms_r Subject: 3D structure of GPCR ligands Date: 1 Feb 1996 05:34:27 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <199602011333.FAA28815@net.bio.net> NNTP-Posting-Host: net.bio.net the crystal structure of human endothelin-1 has been published in: Janes, Peapus, Wallace (1994) Nature Structural Biology 1:311-319 the coordinates have been deposited in the Brookhaven Protein Data Bank as entry 1edn.pdb. alternatively, i can email them to anyone has trouble accessing them there, if you contact me at ubcg91c@ccs.bbk.ac.uk. cheers bonnie wallace From owner-7tms_r@net.bio.net Wed Jan 31 22:00:00 1996 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer2.itd.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 2D and 3D structure of GPCR ligands Date: Thu, 01 Feb 1996 18:11:24 -0500 Organization: University of Michigan Lines: 30 Message-ID: <3111489C.25C@umich.edu> References: <01I0NQZLRJPKA0URL2@mbcl.rutgers.edu> 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 2.0b6a (Win95; I) WCUI@OCELOT.RUTGERS.EDU wrote: > > Hi fellow 7tmer: > I like to find sources for 2D and 3D structures of GPCR ligands. > Is there already a database that contains such data? I'm not sure that you'll find them listed as such. The general database of protein crystal structures is at: http://www.pdb.bnl.gov By the way did you all see the G protein heterotrimer structures in Dec 15, 1995 Cell and January 26, 1996 Nature? Rick > if so, where are they and how can I get them? > if not, where should I start if I want build such database? > Any suggestions will be fully appreciated. > > Weili Cui, Ph.D. > Senior Scientist > Synaptic Pharmaceutical Corp. -- _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450