From owner-7tms_r@net.bio.net Sun Jan 04 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Tony Harmar Newsgroups: bionet.molbio.proteins.7tms_r Subject: ANNOUNCE: VIP/PACAP mailing list Date: 5 Jan 1998 09:27:28 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 46 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <68r560$irm@net.bio.net> NNTP-Posting-Host: net.bio.net I have created a mailing list for the discussion of research on the regulatory peptides VIP (vasoactive intestinal peptide) and PACAP (pituitary adenylate cyclase activating polypeptide). It is my hope that this list can provide a forum through which scientists interested in VIP and PACAP can exchange information, and discuss topics of general interest. Ultimately, it is hoped that frequently asked questions on this list (e.g. sources of probes and antisera, nomenclature of receptors, laboratories working in the field) together with a searchable archive of postings to the list, will be available on a web page maintained by list members. Postings to the list are informal, and decisions and recommendations formulated there do not constitute any official standards. Neither the administrator of the list nor his employer accept any responsibility for the content of the postings. Please spread this announcement as widely as possible amongst your colleagues. In the early stages, I plan to contact as many people as possible who are involved in this small but important area of research, so to save effort, it would be helpful if you could let me know the names of anyone that you have informed To subscribe to this list: Send email to majordomo@dl.ac.uk with "subscribe vippacap" in the first line of the text of the message To unsubscribe to this list: Send email to majordomo@dl.ac.uk with "unsubscribe vippacap" in the first line of the text of the message Should this fail, if you otherwise need human assistance, or if you wish to comment to the list owner, send a message to: owner-vippacap@dl.ac.uk. To send mail to the list: Send the mail to "vippacap@dl.ac.uk" Further details of the list will be sent to you when you subscribe. The list will become what its users make it: please participate, and please send me your reactions and suggestions for improvement. And be patient: it may take a few weeks for things to gain momentum. - ----------------------------------------------------------------------- Tony Harmar Tony.Harmar@ed.ac.uk MRC Brain Metabolism Unit Royal Edinburgh Hospital Morningside Park Tel +44 131 537 6533 Edinburgh EH10 5HF, Scotland FAX +44 131 537 6110 http://sirius.lns.ed.ac.uk/community/members/harmar.html From owner-7tms_r@net.bio.net Tue Jan 06 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Hind van Tol Steye Newsgroups: bionet.molbio.proteins.7tms_r Subject: beta-gamma subunits Date: 7 Jan 1998 12:17:29 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <690nsp$6vj@net.bio.net> Reply-To: Hind van Tol Steye NNTP-Posting-Host: net.bio.net Dear netters, I would like to do some experiments in which G-protein beta-gamma subunits should be injected into cells, to see whether this causes any changes in potassium- or calcium currents. Does anybody know where we can get beta-gamma subunits, as we are not able to produce/purify them ourselves? Thanks, Hind van Tol. From owner-7tms_r@net.bio.net Tue Jan 13 22:00:00 1998 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.molbio.proteins.7tms_r Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 14 Jan 1998 11:46:50 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 232 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <69j4na$g8m@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! 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. From owner-7tms_r@net.bio.net Tue Jan 13 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Athan Kuliopulos Newsgroups: bionet.molbio.proteins.7tms_r Subject: Signal Transduction Postdoc Available/Boston Date: 14 Jan 1998 11:47:18 -0800 Organization: NEMC Lines: 52 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <69j4o6$gaq@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 Thu Jan 15 22:00:00 1998 Path: biosci!biosci!not-for-mail From: forte@ohsu.EDU (Michael Forte) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Two Hybrid Date: 16 Jan 1998 11:27:34 -0800 Organization: Vollum Institute Lines: 9 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <69ocb6$6dj@net.bio.net> NNTP-Posting-Host: net.bio.net Hi, I know this has been discussed here in the past but, to refresh my memory, perhaps someone can recount experiences trying to use 7TM receptors in two hybrid screens. Someone here is trying to convince me that this makes sense, expecially now that activating mutations for 7TMs have been found. I remain skeptical. Mike Forte From owner-7tms_r@net.bio.net Sun Jan 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: pingw Newsgroups: bionet.molbio.proteins.7tms_r Subject: Postdoc at UCSF Date: 19 Jan 1998 16:31:18 -0800 Organization: UCSF Lines: 16 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6a0r8m$aga@net.bio.net> Reply-To: pingw@itsa.ucsf.edu NNTP-Posting-Host: net.bio.net A postdoctoral fellowship position opens immediately in the Division of Immunology/Allergy of Department of Medicine and Immunology/Microbiology at University of California-San Francisco (Parnassus Campus). The group is studying the newly-identified G protein-coupled receptors for the recently-recognized phospholipid growth factors, such as lysophosphatidic acid (LPA) and sphingolipids. Several cDNAs for these receptors have been just cloned and identified in this lab, and studies on genetic, biochemical, pharmacological and functional characterization of these cloned receptors are being actively carried out. We seek motivated candidates with a recent Ph.D. or MD/Ph.D. degree and research experience in protein biochemistry, and basic molecular and cellular biology. Interest in various aspects of G protein-coupled receptors is desired. Stipend starts from $30,000 and depends on qualification. Send CV and names of references to Dr. Edward J. Goetzl, Department of Medicine and Immunology/Microbiology, UCSF, Box 0711, San Francisco, CA 94143-0711, or reply to this email address. songzh@itsa.ucsf.edu From owner-7tms_r@net.bio.net Mon Jan 19 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Cornelius Krasel Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Two Hybrid Date: 20 Jan 1998 06:02:22 -0800 Organization: University of Wuerzburg, Germany Lines: 25 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6a2ape$b2k@net.bio.net> References: <69ocb6$6dj@net.bio.net> NNTP-Posting-Host: net.bio.net Michael Forte wrote: > I know this has been discussed here in the past but, to refresh my memory, > perhaps someone can recount experiences trying to use 7TM receptors in two > hybrid screens. Someone here is trying to convince me that this makes > sense, expecially now that activating mutations for 7TMs have been found. > I remain skeptical. The only published two hybrid paper I know of was done by the group of Mark von Zastrow (UCSF). They screened a two-hybrid library with a sequence from the beta-2-adrenergic receptor and found interaction with the alpha subunit of initiation factor 2B: U. Klein, M. T. Ramirez, B. K. Kobilka, M. von Zastrow (1997): A novel interaction between adrenergic receptors and the alpha subunit of eukaryotic initiation factor 2B. J. Biol. Chem. 272, 19099-19102. As far as I recall (but you'd better go to the library yourself :-), they did not find interactions to G-proteins or arrestins. - --Cornelius. - -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Tue Jan 20 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Vanessa Le Newsgroups: bionet.molbio.proteins.7tms_r Subject: Microvessel endothelial cell search Date: 21 Jan 1998 09:19:45 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6a5anh$pos@net.bio.net> NNTP-Posting-Host: net.bio.net Hello, I am a graduate student in Pharmaceutical Sciences from UT @ Austin. I am currently in search of a capillary endothelial cell line (could be isolated from any species) to develop an in vitro BBB model system to study drug transport. I have contacted various research labs as well as several cell culture distributors, but I still have not been successful in acquiring a batch of this cell culture. I would like to thank in advance anyone who may be able to help me out with this or perhaps know of someone who has accessed to this cell line. Vanessa Le From owner-7tms_r@net.bio.net Wed Jan 21 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Christophe GUENIN" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Dimer Date: 22 Jan 1998 12:39:08 -0800 Organization: None Lines: 10 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6a8apc$m6t@net.bio.net> NNTP-Posting-Host: net.bio.net I have a dimerisation of receptor-G protein ( 7TM-7TM and 7TM-6TM). Does anybody know the phenomenon ? Thanks to give me explanation and bibliographic references. Sophie Paris,France e mail : christophe.guenin@hol.fr From owner-7tms_r@net.bio.net Fri Jan 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: John McLean Newsgroups: bionet.molbio.proteins.7tms_r Subject: CREB with mutated KID domain Date: 24 Jan 1998 10:07:11 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 25 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6adakf$b4p@net.bio.net> NNTP-Posting-Host: net.bio.net We willl have very shortly have finished making a series of GST- CREB fusion proteins with mutated KID domain. We are attempting to make CREB a better substrate G kinase. We have changed two arginines to lysines (N terminal to phosphorylated serine), the phosphorylated serine to threonine and the C terminal isoleucine to a phenyalanine. This was done in a stepwise manner so we have a number of variants. The literature suggests these changes may make a CREB a better substrate for G kinase. We have peptide data that suggests this may be the case, Would anyone like access to the fusion proteins. They may be suitable for persons working on the role of the KID domain in binding to mleocules such as CBP. I have only a small research group and wish to get as much mileage out of the mutated proteins as possible as it represented a fair amount of work for us. Anyone interested ? Any use to anyone ? Dr. John McLean Biological Sciences University of Paisley Scotland From owner-7tms_r@net.bio.net Mon Jan 26 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Ian Murray" Newsgroups: bionet.molbio.proteins.7tms_r Subject: 7TM and gels Date: 27 Jan 1998 10:04:33 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6al7jh$d14@net.bio.net> NNTP-Posting-Host: net.bio.net Hello, I am trying to run a 7TM crosslinked with radiolabelled ligand on gels for visualisation of their interaction. We are using isolated cell membranes for binding with separation of free ligand via vacuum filtration through a filter membrane. The filter membrane is isolated and heated at 60C with 1X SDS loading buffer. When this is run on a 10% polyacrylamide gel, a band is seen at the interface of the stacking and separating gels (and another for free ligand). However, when run on a 5% acrylamide/PDA gel (without SDS) a band is seen at 75000MW. Does anyone have an explanation for this, is it due to the interface between the gels, pH (separating and stacking) or lack of SDS in the PDA gel or aggregation?. Furthermore, does anyone have a gel system that is reliable and avoids this problem. (I will try a gradient gel tommorow). From owner-7tms_r@net.bio.net Mon Jan 26 22:00:00 1998 Path: biosci!biosci!not-for-mail From: newsmgr@merrimack.edu Newsgroups: bionet.molbio.proteins.7tms_r Subject: US-NY-PROTEIN PURIFICATION POSITION Date: 27 Jan 1998 10:04:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6al7k4$d7f@net.bio.net> NNTP-Posting-Host: net.bio.net Relay-Version: ANU News - V6.2.0 06/23/97 OpenVMS AXP V6.2; site chasm Path: chasm!cam-news-feed2.bbnplanet.com!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!news.idt.net!ix.netcom.com!news Newsgroups: bionet.molbio.proteins.7tms_r Subject: US-NY-PROTEIN PURIFICATION POSITION Message-ID: <01bd2b29$4d3def40$2f79d6ce@scott-s-armada> From: "Scott Shanes" Date: 27 Jan 1998 13:40:44 GMT Organization: Netcom Lines: 15 NNTP-Posting-Host: trn-nj1-15.ix.netcom.com X-NETCOM-Date: Tue Jan 27 7:40:44 AM CST 1998 X-Newsreader: Microsoft Internet News 4.70.1161 I am looking for a person to perform protein purification. The candidate will have 1+ years experience with protein purification (general chromatography skills) and characterization (SDS Page, Western Blot, slot/dot blot, HPLC). Familiarity with UV VIS spectroscopy and computers. You will be responsible for the purification of proteins in small to medium scale from baculovirus infected cells and mammalian cells and purification and refolding of proteins from E. coli and Pichia sources. You should possess a B.S. or M.S.degree in Biochemistry or medical sciences. We are a leading biotech firm with research facilities in Tarrytown, New York and can provide excellent benefits (health insurance, dental, and vision plan, paid vactation and more). A high impact, high profile position with excellent opportunity for advancement. Please contact Scott Shanes by phone at 609-584-8733 Ext. 218, fax resume and cover letter to 609-584-9575 or E-Mail to sis@diedremoire.com From owner-7tms_r@net.bio.net Wed Jan 28 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Cornelius Krasel Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Two Hybrid Date: 29 Jan 1998 21:23:33 -0800 Organization: University of Wuerzburg, Germany Lines: 14 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6aro4l$s7h@net.bio.net> References: <69ocb6$6dj@net.bio.net> <6a2ape$b2k@net.bio.net> Reply-To: hadcocj@war.wyeth.com NNTP-Posting-Host: net.bio.net [I received an email by John Hadcock which apparently was also intended for the group but may have become lost; at least it has not appeared on my server yet. Reply-To: set. -- Cornelius.] Hi all, Brad Ozenberger (Kajkowski et al) demonstrated a 2-hybrid interaction between the N-terminus of the GHRH receptor and GHRH (J. of Receptor and Signal Transduction Research 17(1-3):293-303, 1997). A concern that I have on 2-hybrid is the "pharmacological integrity" of the 2-hybrid system for examining these interactions. Many of the interaction are low affinity and need to be confirmed in mammalian expression systems. John Hadcock hadcocj@war.wyeth.com From owner-7tms_r@net.bio.net Wed Jan 28 22:00:00 1998 Path: biosci!biosci!not-for-mail From: newsmgr@merrimack.edu Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Dimer Date: 29 Jan 1998 21:23:51 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 52 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6aro57$s82@net.bio.net> NNTP-Posting-Host: net.bio.net Relay-Version: ANU News - V6.2.0 06/23/97 OpenVMS AXP V6.2; site chasm Path: chasm!cam-news-feed2.bbnplanet.com!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!news.idt.net!peerfeed.ncal.verio.net!newsfeed.slip.net!newsfeed.interlog.com!news.interlog.com!yblee Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Dimer Message-ID: <6ad761$nsi$1@news.interlog.com> From: samuel.lee@utoronto.ca (Samuel P. Lee) Date: Sat, 24 Jan 98 17:18:44 GMT References: <6a8apc$m6t@net.bio.net> Organization: University of Toronto Lines: 39 NNTP-Posting-Host: ip226-6.cc.interlog.com X-Newsreader: News Xpress Version 1.0 Beta #4 In article <6a8apc$m6t@net.bio.net>, "Christophe GUENIN" wrote: >I have a dimerisation of receptor-G protein ( 7TM-7TM and 7TM-6TM). > >Does anybody know the phenomenon ? > >Thanks to give me explanation and bibliographic references. > >Sophie >Paris,France > >e mail : christophe.guenin@hol.fr If you mean dimerization of two receptors, there many research groups (including our lab) that are studying this phenomenon. It had been suspected for a long time that G protein coupled receptors may form dimers. In the past few years, there have been several papers that deal with this issue directly. Here is a list of some publications (It is definitely not a comprehensive list, but, it's a start): Maggio et al., PNAS 90: 3103, 1993 Ng et al., Biochem. Biophys. Res. Comm. 227: 200, 1996 Hebert et al., J. Biol. Chem. 271: 16384, 1996 Fukushima et al., FEBS Lett. 409: 283, 1997 Cvejic and Devi, J. Biol. Chem., 272: 26959, 1997 Nimchinsky et al., J. Biol. Chem., 272: 29229, 1997 I hope this helps. Sam Lee samuel.lee@utoronto.ca Department of Pharmacology University of Toronto Toronto, ON, Canada M5S 1A8 From owner-7tms_r@net.bio.net Wed Jan 28 22:00:00 1998 Path: biosci!biosci!not-for-mail From: newsmgr@merrimack.edu Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Two Hybrid Date: 29 Jan 1998 21:24:04 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 54 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6aro5k$sbc@net.bio.net> NNTP-Posting-Host: net.bio.net Relay-Version: ANU News - V6.2.0 06/23/97 OpenVMS AXP V6.2; site chasm Path: chasm!cam-news-feed2.bbnplanet.com!cam-news-hub1.bbnplanet.com!news.bbnplanet.com!news.idt.net!peerfeed.ncal.verio.net!newsfeed.slip.net!newsfeed.interlog.com!news.interlog.com!yblee Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Two Hybrid Message-ID: <6ad7l3$nsi$2@news.interlog.com> From: samuel.lee@utoronto.ca (Samuel P. Lee) Date: Sat, 24 Jan 98 17:26:46 GMT References: <69ocb6$6dj@net.bio.net> <6a2ape$b2k@net.bio.net> Organization: University of Toronto Lines: 42 NNTP-Posting-Host: ip226-6.cc.interlog.com X-Newsreader: News Xpress Version 1.0 Beta #4 I do not know the specific reference, but H. Van Tol's group at the University of Toronto has used the yeast 2 hybrid screen with segments of the D4 Dopamine receptor. I believe the group was examining interactions with the repeat sequence in the third IC loop. Sam Lee samuel.lee@utoronto.ca Department of Pharmacology University of Toronto Toronto, ON, Canada M5S 1A8 In article <6a2ape$b2k@net.bio.net>, Cornelius Krasel wrote: >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. > >The only published two hybrid paper I know of was done by the group of >Mark von Zastrow (UCSF). They screened a two-hybrid library with a >sequence from the beta-2-adrenergic receptor and found interaction >with the alpha subunit of initiation factor 2B: > >U. Klein, M. T. Ramirez, B. K. Kobilka, M. von Zastrow (1997): >A novel interaction between adrenergic receptors and the alpha subunit >of eukaryotic initiation factor 2B. J. Biol. Chem. 272, 19099-19102. > >As far as I recall (but you'd better go to the library yourself :-), they >did not find interactions to G-proteins or arrestins. > >- --Cornelius. > >- -- >/* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ >/* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ >/* "Science is the game we play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Wed Jan 28 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Gert.Vriend@EMBL-Heidelberg.de Newsgroups: bionet.molbio.proteins.7tms_r Subject: 7tm models Date: 29 Jan 1998 21:33:23 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6aron3$8k@net.bio.net> NNTP-Posting-Host: net.bio.net Dear 7-helical friends, We have built models of 650 Class A GPCRs based on the helix orientations and alpha carbon positions that are described in: Baldwin J.M., Schertler G.F.X., Unger V.M. J. Mol. Biol., 272 (1997) 144-164 An alpha-carbon template for the transmembrane helices in the rhodopsin family of G-protein-coupled receptors. These coordinates were kindly made available to us by J.Baldwin. These models are available via the GPCRDB. Be aware though that these models were built in a 100% automatic mode. There are just too many models to check them all by hand. So, if you really are going to use one of these models in your own research, please get help from a specialist if needed and check the model carefully. If you find 'errors' please report them so we can fix things. The addresses are: GPCRDB: http://www.gpcr.org/7tm/ The model page: http://www.gpcr.org/7tm/models/ The models mentioned above: http://www.gpcr.org/7tm/models/vriend2/ Explanation/disclaimer/acknowledgements: http://www.gpcr.org/7tm/models/vriend/modelinf.html Yours, the GPCRDB staff. From owner-7tms_r@net.bio.net Sat Jan 31 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Cynthia S. Smagula" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Free 1998 Source Book Available to Researchers Date: 1 Feb 1998 10:44:39 -0800 Organization: BIOTA Publications Lines: 21 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6b2fqn$1pv@net.bio.net> Reply-To: biota@onramp.net NNTP-Posting-Host: net.bio.net Obtain a free copy of the 1998 Source Book, a comprehensive guide to bioresearch products, by filling in the request form at the BioSupplyNet site: http://www.biosupplynet.com. The Source Book provides up-to-date information about molecular biology products, lab instrumentation, and supplies. 70,000 copies of the Source Book will be given to scientists this year. Biomedical researchers will find that the unique organization of the Source Book simplifies product searching and speeds the gathering of comparative information. The Source Book indexes products in 2250 categories, and provides up-to-date contact information, including WWW and e-mail addresses, for 3000 suppliers. Bench Basics comparative tables allow apples-to-apples comparisons of products. A special Lab Set-up Check List simplifies the process of putting together a new laboratory. The BioSupplyNet web site at http://www.biosupplynet.com ,the online companion to the Source Book, provides interactive access to Source Book information as well as several enhancements, including the BioToolKit, a searchable directory of 500 free molecular biology databases, analytic tools, and structural imaging software applications. From owner-7tms_r@net.bio.net Sat Jan 31 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Gert.Vriend@EMBL-Heidelberg.de Newsgroups: bionet.molbio.proteins.7tms_r Subject: GPCR Models ay GPCRDB Date: 1 Feb 1998 10:45:57 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6b2ft5$2il@net.bio.net> NNTP-Posting-Host: net.bio.net 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 -------