From owner-7tms_r@net.bio.net Mon Feb 02 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Fabien Campagne Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: GPCR Models ay GPCRDB Date: 3 Feb 1998 16:14:57 -0800 Organization: Lab. Chimie theorique Nancy Lines: 35 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6b8bu1$46r@net.bio.net> References: <6b2ft5$2il@net.bio.net> NNTP-Posting-Host: net.bio.net Hi Gert, I saw your message in the bionet.molbio.proteins.7tms_r newsgroup Subject: 7tm models Date: 29 Jan 1998 21:33:23 -0800 There were a few messages on the group since the beginning of the year see you in march, Fabien Gert.Vriend@EMBL-Heidelberg.de wrote: > Dear 7tm friends, > > Ten days ago I posted the message that we have built about > 650 GPCR models based on the coordinates that Joyce Baldwin > created from a combination of sequence analyses and looking > at the low resolution electron density of rhodopsin. > > Neither did I receive this mail myself, nor did I get any > questions about the models, so I guess the mail got lost > (actually I did not get any messages from this group for > several months now...). > > Therefore again, if you are interested in these new models, > point your web-browser at: > http://www.gpcr.org/7tm/ > and go to the models page. > > Greetings in sevenfold > > Gert Vriend > ------- End of forwarded message ------- From owner-7tms_r@net.bio.net Mon Feb 02 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Athan Kuliopulos Newsgroups: bionet.molbio.proteins.7tms_r Subject: Signal Transduction Postdoc Avail/Boston Date: 3 Feb 1998 16:14:59 -0800 Organization: NEMC Lines: 52 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6b8bu3$474@net.bio.net> Reply-To: "Kuliopoulos@HemOnc"@NEMC.nemc.org NNTP-Posting-Host: net.bio.net Postdoctoral Position Available Center of Hemostasis and Thrombosis Research Tufts University School of Medicine-NEMC Boston, MA We have an immediate opening for a Postdoctoral Fellow to work on thrombin receptor-dependent signal transduction. The discovery of a thrombin receptor that is activated by proteolytic cleavage and exposure of a new N-terminal tethered ligand has generated considerable interest because it is the first example of a protease-activated membrane receptor. Receptor activation precipitates complex signaling events culminating in platelet aggregation, wound healing, and cellular proliferation. Since chronic activation of the receptor may lead to coronary artery disease, stroke, and other vascular diseases, preventing thrombin receptor activation is of pharmacologic interest. The project involves genetic and biochemical studies of the thrombin receptor expressed in yeast. Heterologous expression of a thrombin receptor-G-protein pair will activate a defined signal transduction pathway that leads to a distinct, unambiguous phenotype=97growth. We hav= e recently produced and purified affinity-tagged thrombin receptor in yeast which can be cleaved by nanomolar thrombin. The receptor couples to specific mammalian G proteins, however, the molecular basis for the coupling specificity is unknown. Genetic studies of receptor, G proteins, ligand, and downstream regulators will greatly aid in understanding how this receptor activates mammalian signaling pathways. The yeast expression system of the receptor coupled to the endogenous yeast MAP kinases may provide a screening method for anti-thrombin receptor drugs. Our laboratory is located within the Center of Hemostasis and Thrombosis Research, a modern, well-equipped facility supported by several core units including automated DNA sequencing, oligonucleotide and peptide synthesis, plasmid preparation, fermentation, and flow cytometry. Qualifications for this position are a Ph.D. degree and US citizenship or permanent residency. Candidates with training in yeast genetics who would like to acquire expertise in biochemistry in the context of cardiovascular disease are especially encouraged to apply. Interested candidates should e-mail a description of their research interests, a CV, and names of three references to: Athan Kuliopulos, MD., Ph.D. Assistant Professor Departments of Medicine and Biochemistry Tufts-NEMC Box 832 750 Washington Street Boston, MA 02111 617-636-8482 617-636-4833 (fax) akuliopu@opal.tufts.edu From owner-7tms_r@net.bio.net Wed Feb 04 22:00:00 1998 Path: biosci!biosci!not-for-mail From: vheerik@chem.vu.nl (Harm van Heerikhuizen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re:Two Hybrid Date: 5 Feb 1998 10:54:17 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 54 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6bd1sp$m8r@net.bio.net> NNTP-Posting-Host: net.bio.net Dear Michael, Michael Forte wrote: > I know this has been discussed here in the past but, to refresh my memory, > perhaps someone can recount experiences trying to use 7TM receptors in two > hybrid screens. Someone here is trying to convince me that this makes > sense, expecially now that activating mutations for 7TMs have been found. > I remain skeptical. I recently saw a paper describing the reverse 2-hybrid. That is, using a G-protein alpha subunit as a bait to find a GPCR. This may suggest that your scepticism is not justified. The reference is: Yun CW. Tamaki H. Nakayama R. Yamamoto K. Kumagai H. G-PROTEIN COUPLED RECEPTOR FROM YEAST SACCHAROMYCES CEREVISIAE. Biochemical & Biophysical Research Communications. 240(2):287-292, 1997 Nov 17. Abstract: The Saccharomyces cerevisiae GPR1 (G-protein coupled receptor) gene was isolated using two-hybrid system with a heterotrimeric GTP binding protein alpha subunit Gpa2p as a bait. The GPR1 gene encodes 961 amino acids with predicted seven transmembrane segments and two large cytosolic regions as third cytosolic loop with 350 amino acids where asparagine-rich region was found and the C-terminal region with 283 amino acids. The Gpr1p interacted with Gpa2p at C-terminal region with 131 amino acid residues as well as third cytosolic loop. Disruption of the GPR1 gene was not lethal and did not affect to the cell growth. The Gpr1p-GFP fusion protein localized at the cell surface. These results suggest that Gpr1p is a G-protein coupled receptor which localized at plasma membrane, It is likely that a Gpr1p monitors the extracellular signal such as nutrition and transduce it via Gpa2p a possible positive regulator of cAMP level. Good luck with your screens! Harm *************************************************************************** Harm van Heerikhuizen, PhD Dept. of Biochemistry and Molecular Biology Vrije Universiteit De Boelelaan 1083 1081 HV Amsterdam The Netherlands tel: + 31 - 20 - 444 7573 (office) tel: + 31 - 20 - 444 7555 (secretary) tel: + 31 - 20 - 672 2160 (home) fax: + 31 - 20 - 444 7553 e-mail: vheerik@chem.vu.nl *************************************************************************** From owner-7tms_r@net.bio.net Thu Feb 12 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Stuart Sealfon" Newsgroups: bionet.molbio.proteins.7tms_r Subject: inducible expression Date: 13 Feb 1998 08:24:23 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6c1s3n$1de@net.bio.net> NNTP-Posting-Host: net.bio.net Has anyone had experience with the clontech Tet-On system or the Invitrogen ecdysone system for driving GPCR expression in cell lines? We have tried both systems for the GnRH receptor and have not gotten any detectable binding. The luciferase or beta-gal control works in both, but not very impressively. I'm curious if this is a specific receptor problem or if these systems just aren't strong enough promoters when induced for receptor expression. Stuart Sealfon From owner-7tms_r@net.bio.net Thu Feb 12 22:00:00 1998 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molbio.proteins.7tms_r Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 13 Feb 1998 08:24:31 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 232 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6c1s3v$1ed@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-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/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 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. 0) BIOSCI NEEDS YOUR SUPPORT!! - ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 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. 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! 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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. From owner-7tms_r@net.bio.net Mon Feb 16 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Ellen R. Weiss" Newsgroups: bionet.molbio.proteins.7tms_r Subject: phosphopeptides Date: 17 Feb 1998 04:21:16 -0800 Organization: University of North Carolina Lines: 6 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cbvbs$qk0@net.bio.net> Reply-To: erweiss@med.unc.edu NNTP-Posting-Host: net.bio.net Does anyone know a good, reputable place to order synthetic peptides phosphorylated on serines and threonines? Thanks, Ellen Weiss From owner-7tms_r@net.bio.net Mon Feb 16 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Jonathan Lee Newsgroups: bionet.molbio.proteins.7tms_r Subject: JURKAT ENDOGENOUS GPCRS Date: 17 Feb 1998 04:21:17 -0800 Organization: Eli Lilly and Company Lines: 22 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cbvbt$qk8@net.bio.net> NNTP-Posting-Host: net.bio.net Dear 7TMers, I believe there was a some discussion about endogenous GPCRs in Jurkat cells a few months ago, but I have not been able to find any information. Does anyone have any references or information??? Thanks. Dr. Jonathan A. Lee Research Technologies and Protein Mail Stop 1533 Eli Lilly and Company Lilly Corporate Center Indianapolis, Indiana 46285 Phone: 317-277-8123 Fax: 317-276-5281 EMAIL: Jonathan_A_Lee@lilly.com From owner-7tms_r@net.bio.net Mon Feb 16 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Ilya Vakser Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdocs/Graduate Students in Protein Docking and Structure Prediction Date: 17 Feb 1998 04:21:19 -0800 Organization: Medical University of South Carolina Lines: 35 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cbvbv$qkk@net.bio.net> NNTP-Posting-Host: net.bio.net Applications are invited for postdoctoral and graduate student positions in my laboratory at the Medical University of South Carolina. The positions were advertised in November, however, due to the delayed release of funds, I post them again. The main subjects of the research in the laboratory are computational studies of molecular recognition, docking methodology, structure prediction, and applications to signal transduction pathways and other molecular systems. For more information, see the lab's web site at http://reco3.musc.edu. The following positions are immediately available: Postdoctoral fellow Development of flexible docking techniques for small ligands. Docking methodology for computer-aided drug design. Postdoctoral fellow Modeling of integral membrane receptors. Structure prediction based on the principles of molecular recognition. Graduate student General development of protein docking methodology. Strong programming skills in C are required for all positions. To apply send or email a letter and CV with names of 3 referees. Ilya A. Vakser Assistant Professor of Pharmacology Department of Cell and Molecular Pharmacology Medical University of South Carolina 171 Ashley Avenue Charleston, SC 29425 Email: vakseri@musc.edu, Phone:(803)792-2471, Fax:(803)792-2475 From owner-7tms_r@net.bio.net Tue Feb 17 22:00:00 1998 Path: biosci!biosci!not-for-mail From: roth@biochemistry.BIOC.CWRU.Edu (B.L. Roth) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Post-doc position Date: 18 Feb 1998 12:28:51 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 19 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cfga3$7qf@net.bio.net> NNTP-Posting-Host: net.bio.net A postdoctoral position is available in June to study molecular mechanisms by which GPCR's couple to G proteins and accessory docking proteins. Background in molecular biology is required. The position is firm for two or three years. For a description of lab and current projects, please visit my web site which is listed below. BL Roth MD, PhD Department of Biochemistry Room W438, Wood Building 10900 Euclid Avenue Case Western Reserve University Medical School Cleveland, OH 44106-4935 http://www.freeyellow.com/members/blroth 216-368-2370 (Office) 216-368-3419 (Fax) roth@biocserver.cwru.edu (email) From owner-7tms_r@net.bio.net Wed Feb 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: TJ Murphy Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoc: Vascular Gene Expression Date: 19 Feb 1998 08:30:20 -0800 Organization: Emory University Lines: 85 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6chmms$n25@net.bio.net> NNTP-Posting-Host: net.bio.net Postdoctoral Position Available Regulation of Vascular Gene Expression Emory University I am looking for a postdoctoral fellow to study receptor-mediated control of gene expression. The position is available as early as May of 1998. General interests in the lab largely revolve around receptors, signaling, gene expression and vascular biology. Our model systems include studies of gene expression control in both vascular smooth muscle and endothelial cells, and very recently in transgenic mice. Two projects are under study. One involves the control of mRNA stability and translation by receptor-mediated signaling. The second project involves the control of a heteromeric transcription factor complex by cell surface receptors. Activation of this complex leads to highly inducible transcription of target genes in both vascular smooth muscle and endothelial cells. Since each of these systems are activated by multiple classes of receptors, they both serve as experimentally approachable models for solving the general mechanisms of how the many complex signals emanating from cell surface receptors are filtered to achieve specific responses. Also, we have data predicting the existence of unknown but key effector components in each of these systems, which we will need to isolate/clone to gain better insight into their mechanisms. A third issue revolves around understanding the role of these gene expression systems in regulating a few candidate target genes we have identified recently, using both cell culture and animal models. The postdoctoral fellow would be expected to focus initially on a single aspect of any one of these issues as appropriate to their own interests and talents. We have recently developed some novel and highly efficient retroviral vectors and protocols to express foreign genes in primary cultures of vascular cells. These include several types of transcriptional reporters, fusion proteins that can function as cellular antagonists of signaling enzymes and transcription factors, and precisely specified mutant mRNA's under selective transcriptional regulation. The lab exploits a range of cell and molecular methodologies. Wider use of transgenic mouse models is contemplated in the very near future, including targeted expression of recombinant genes to the vasculature of mice to inhibit signaling and transcription selectively in vivo. This is a highly attractive opportunity that would allow for an energetic individual to become immediately successful using our well-established models and approaches. There are also several conceivable exit pathways, in the form of attractive problems related to these interests that are we are unlikely to pursue over the long term. A successful candidate can begin to develop these in my lab as a basis to start their own independent career. The ideal candidate will be a recent graduate with a strong record of publication. An interest and familiarity with either vascular biology, receptors and signal transduction, or regulated gene expression is essential. I am particularly interested in identifying individuals who have a solid background in experimental cardiovascular physiology and wish to continue along those lines, but who would like to learn how to manipulate gene expression in vivo through exploitation of transgenic technology. The Pharmacology Department at Emory is thriving with a highly collegial, 14 member faculty, which is involved in training over 40 postdocs and students. Sufficient funding is available in my own lab to accomodate the needs of 6-8 scientists. Our interdisciplinary graduate training programs foster interactions among various Departments and Centers in the Emory biomedical community. The University is located in an attractive, affordable and close-in suburban community, and is near many cultural and recreational venues in the southeast region of the US. Please contact me if you are interested, or pass this note along to someone who might be. Interested individuals should send along a CV and reprints or your work. Thanks - -- T.J. Murphy, Ph.D. 404-727-2467 Assistant Professor 404-727-0365 (fax) Department of Pharmacology tmurphy@pharm.emory.edu Emory University School of Medicine 5031 O.W. Rollins Research Building Atlanta, GA 30322 http://www.emory.edu/PHARMACOLOGY/MURPHY/ "If we're not driving paradigms, then we're out driving Titleists" From owner-7tms_r@net.bio.net Mon Feb 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: rappaport@email.chop.edu (Eric Rappaport) Newsgroups: bionet.molbio.proteins.7tms_r Subject: DNA Sequencers for Sale Date: 24 Feb 1998 09:15:13 -0800 Organization: Children's Hospital of Philadelphia Lines: 69 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cuv71$cp@net.bio.net> NNTP-Posting-Host: net.bio.net AUTOMATED DNA SEQUENCERS FOR SALE Three ABI 373 DNA sequencers are available for sale. Please contact Dr. Eric Rappaport (rappaport@email.chop.edu) with offers or questions. Details of the equipment configuration follow: Two 373 units with Stretch upgrades: Filters - 5-filter wheel (can choose between 2 different sets of 4 different dyes). Chemistries - Standard Taq (FS) terminator, dye primer or ABI T7 DNA polymerase sequencing chemistry. All current Genescan chemistries, including microsatellite panels, primers labeled during synthesis with standard ABI dyes (6-Fam, Hex, Tet, Tamra) or (6-Fam, Hex, Tamra, and Rox). Gel lengths - Multiple gel (well-to-read) lengths including 48 cm (standard long-read [>650 nt] sequencing length), 34 (shorter sequence reads [550-600 nt] or high-resolution Genescan applications), 24 (microsatellite applications) 12 (lower-resolution Genescan applications) or 6 (lower-resolution Genescan applications). Scan Speed - 1X, BaseSprinter (2X, but limited to middle half of gel; can be used for faster analysis of shorter sequences). One 373 with multiple gel length (Leon) configuration: Filters - 5-filter wheel (can choose between 2 different sets of 4 different dyes). Chemistries - Standard Taq (FS) terminator, dye primer or ABI T7 DNA polymerase sequencing chemistry. All current Genescan chemistries, including microsatellite panels, primers labeled during synthesis with standard ABI dyes (6-Fam, Hex, Tet, Tamra) or (6-Fam, Hex, Tamra, and Rox). Gel lengths - Multiple gel (well-to-read) lengths including 24 cm (shorter sequence reads [400-500 nt] or microsatellite applications) 12 (lower-resolution Genescan applications) or 6 (lower-resolution Genescan applications). Scan Speed - 1X, BaseSprinter (2X, but limited to middle half of gel; can be used for faster analysis of shorter sequences). Computers and Software Unit 1 (Stretch) - Mac IIcx with 8 mB RAM/230 mB hard drive Unit 2 (Stretch) - Mac IIci with 8 mB RAM/100 mB hard drive Unit 3 (Leon) - Mac IIci with 8 mB RAM/120 mB hard drive All units are equipped with latest versions of analysis and collection sofware for running sequencing and Genescan applications on the above computers. Upgrades for PowerMac are not included. Service Contracts All units have been under service contract since the time of their purchase from ABI and have onlt recently been removed from those agreements. Purchaser would need to have an installation service call from ABI before initiation of a new service contract could occur. Shipping Purchaser is responsible for all shipping charges. ABI will sell shipping crates which are appropriate for the units, if necessary. From owner-7tms_r@net.bio.net Mon Feb 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Michal Opas Newsgroups: bionet.molbio.proteins.7tms_r Subject: Calreticulin Workshop: Program & Gen Info Date: 24 Feb 1998 09:17:30 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 212 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6cuvba$1db@net.bio.net> NNTP-Posting-Host: net.bio.net Dear Colleague, We are delighted to announce that Calreticulin Workshop, devoted to the structure and function of calreticulin and related proteins, will take place on March 31 - April 2, 1998 in Banff, Alberta, Canada. The Calreticulin Workshop is a satellite meeting to the 8th Fisher Winternational Symposium on Cellular and Molecular Biology which will be held April 2-5, 1998, also at the Banff Conference Centre. The Workshop will provide unique opportunity to meet and interact with the scientists interested in calreticulin research in spectacular surroundings of Banff National Park in Canadian Rocky Mountains. We are sure that the Banff Calreticulin Workshop will be an important forum to share the latest findings and to develop future interactions. Calreticulin has been implicated to play a role in almost every aspect of cell biology as outlined in a brief overview below. We hope that the Workshop will be useful to sort out some of the latest discoveries and controversies concerning calreticulin and implication of this protein in a variety of biological systems. On the behalf of the Organizing Committee we would like to invite you to participate in the Workshop. Program of the 3rd Calreticulin Workshop "Wine & cheese" reception: March 31, 1998 8:00-10:30 pm Day 1 April 1 9:00-9:20 M. Opas Foreword ER Signalling 9:20-10:00 K. Mikoshiba "Molecular Dynamics in Brain Development" Calcium Chair: N.S. Allen 10:00-10:40 T. Pozzan "Calreticulin as a Ca2+-buffer in the ER: evidence from overexpression studies" 10:40-11:20 KH. Krause "Ligand-gated and store-operated Ca2+ channels" 11:20-11:40 Coffee Break Calcium continued Chair: K. Mikoshiba 12:00-12:40 P. Camacho "Modulation of intracellular Ca2+ release and uptake by Calreticulin, Calnexin and Calmegin in Xenopus oocytes". 12:40-13:40 Lunch Break ER Chaperones Chair: R. Sontheimer 13:40-14:20 J. Bergeron "ER chaperones" 14:20-15:00 W. Nauseef "Roles of calreticulin and calnexin in ER 'quality control'" 15:00-15:40 D. Hebert "Calreticulin and calnexin: ER chaperone partners" 15:40-16:00 Coffee Break Disease Chair: K-H. Krause 16:00-16:40 R. Sontheimer Calreticulin Autoantibody Detection Employing a Ro Autoantigen-Binding Form of Recombinant Human Calreticulin" 16:40-17:20 P. Eggleton "Dual roles of calreticulin in autoimmunity and innate immunity" 17:20-18:00 B. Ritchie "Calreticulin in Blood Coagulation" 18:00-18:40 G. Needham "Tick Calreticulin" Day 2 April 2nd Gene Chair: T. Pozzan 09:00-09:40 R. Clark "Mechanisms of transcriptional regulation of calreticulin expression" 09:40-10:20 R. St.Arnaud "Specific developmental roles for calreticulin revealed by the knock-out mice" 10:20-11:00 D. Llewellyn "Stress-induced calreticulin expression" 11:00-11:20 Coffee Break Gene continued Chair: P. Eggleton 11:20-12:00 Nakhashi "Calreticulin and RNA processing" Plants 12:00-12:40 Allen/Wyatt/Tsou/Robertson "Calreticulin: a tool to alter cellular calcium in plant cells" 12:40-13:20 Navazio/Mariani "Calreticulin in plant cells" 13:20-14:20 Lunch Break Trafficking Chair: J. Bergeron 14:20-15:00 Johnson "Calreticulin in neuronal cells" Adhesion 15:00-15:40 Murphy-Ullrich "Calreticulin as a mediator of thrombospondin-stimulated focal adhesion disassembly" 15:40-16:00 Coffee Break Adhesion continued Chair: R. Clark 16:00-16:40 Dedhar "Perturbation of integrin mediated adhesion and Ca signalling in calreticulin-deficient cells. 16:40-17:20 Opas "Modulation of cellular adhesiveness by levels of expression of calreticulin" 17:20 Closing discussion >>> Conclusion M. Michalak SCIENTIFIC PROGRAM The scientific sessions for the Workshop will take place on Wednesday, April 1, 1998 from 9:00 AM - circa 7:00 PM. Slide projector, overhead projector, black-board and VHS VCR will be available for your presentations. Presentations: Each speaker will have ~40 minutes for presentation: please make presentation short enough to leave ~10 minutes for discussion. As you can see from the scientific program we are all working in extremely diverse areas of the biological sciences. Therefore, please make sure you devote at least 5 minutes of your talk to introduce the biological system you are studying. There is no need to send abstracts for the Workshop. Posters: Bring one or more posters if you wish! Posters shall be displayed throughout the Workshop, but we shall not formal poster sessions. In keeping with informal manner of the Workshop we expect posters to be on display throughout the duration of the Workshop to provide ample opportunity to discuss them at any time. REGISTRATION & ACCOMMODATION Schedule arrival date is on Tuesday, March 31, 1998. Upon arrival to the Banff Center please register for your accommodation at the Front Office in the foyer of the first floor of Donald Cameron Hall (Administration Building). Check-in time for your room is 2:30 PM on your day of arrival. Check-out time is 12:00 noon on your departure day. Your account may be paid in cash or by credit card (Visa, American Express, Master Card, EnRoute). RECEPTION There will be a reception ("wine & cheese") held for the Workshop participants on Tuesday, March 31, 1998 from 8:00 PM - 10:30 PM. Registration packets for the Workshop and name tags will be available during the reception. WINTERNATIONAL SYMPOSIUM The Calreticulin Workshop is a satellite meeting to the 8th Fisher Winternational Symposium on Cellular and Molecular Biology entitled "Membrane Proteins in Health and Disease" which will be held April 2-5, 1998, also at the Banff Conference Centre. Information about the meetings and registration forms can be obtained by contacting: Dr. Carol E. Cass, Chair Winternational Symposium Department of Oncology University of Alberta Cross Cancer Institute Edmonton, Alberta T6G 1Z2 (403) 432-8320 phone (403) 432-8425 fax email: sherron.becker@cancerboard.ab.ca GROUND TRANSPORTATION Bus service by Brewster Transportation is available directly from Calgary International Airport to Banff at 12:30 PM, 3:00 PM and 6:00 PM. The travel time is about 2 hours and the current one way fare is $28.00. Upon request buses will drop off passengers at the Banff Center. Brewster Transportation numbers are 403-762-6767 and 1-800-661-1152. If you have any further questions or special requirements please contact: Marek Michalak at (403) 492-2256 phone (403) 492-0886 fax E-mail: marek.michalak@ualberta.ca or Michal Opas at: (416) 978-8947 phone (416) 978-3954 fax E-mail: m.opas@utoronto.ca SEE YOU IN BANFF! Cheers Marek Michalak & Michal Opas From owner-7tms_r@net.bio.net Tue Feb 24 22:00:00 1998 Path: biosci!biosci!not-for-mail From: klk@mole.bio.cam.ac.uk (Karen L Kennedy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Post-doc position in Molecular Modelling Date: 25 Feb 1998 09:53:41 -0800 Organization: University of Cambridge, England Lines: 18 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6d1lr5$6fn@net.bio.net> NNTP-Posting-Host: net.bio.net We are urgently seeking a post-doctoral researcher having experience in the three dimensional modelling of proteins. This position is in a group where one of the objectives is to determine the structure and sites of ligand interaction of G-protein coupled receptors and their mechanism of activation. The researcher will work with an expert in this field and other researchers using site-directed mutagenesis and pharmacology. Candidates must not be French nationals.The salary will be in the region of 10,000 FFr per month. Candidates should send a full CV and list of publications as soon as possible to: Dr. D. Fourmy, Bat. L3, CHU Rangueil, 31054 Toulouse, France. Tel.: 05 61 32 24 05 Fax.: 05 61 32 24 03 Email: fourmyd@rangueil.inserm.fr From owner-7tms_r@net.bio.net Thu Feb 26 22:00:00 1998 Path: biosci!biosci!not-for-mail From: John Hadcock Newsgroups: bionet.molbio.proteins.7tms_r Subject: stable expression of GPCRs Date: 27 Feb 1998 12:11:22 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6d76la$4kr@net.bio.net> NNTP-Posting-Host: net.bio.net Hi all, We have encountered a strange problem with regard to the stable expression of a GPCR. We have done transfections using both human and rat homologs (in either pCDNA3 or pRC/CMV or ecdysone-inducible system) in CHO and HEK293 cells. After selection with G418 we pick 12 colonies and test for positives in binding assays. We usually get 200-1000 fmol/mg in binding assays in the initial screen. After 2-3 passages expression levels decline or become inconsistent. As a positive control we use a second subtype. This subtype does not exhibit this strange behavior. We have tried regular serum, dialyzed serum and serum-free medium. The last two seem to help a little in recovering binding but we never see really good expression (as compared to the other receptor subtype). Antagonist binding appears to better than agonist binding and the agonist binding is GTP-sensitive. Has anyone encountered similar problems? Any comments and suggestions would be helpful and appreciated. Also, does anyone know of a supplier of Muristerone A. Invitrogen has sold a wonderful ecdysone-inducible system. However, their supplier of muristerone A no longer sells it. Thus, they sell a system that is non-functional without the inducer. Thank you, John Hadcock Wyeth-Ayerst Research hadcocj@war.wyeth.com