From owner-7tms_r@net.bio.net Tue Oct 08 23:00:00 1996 Path: biosci!FRODO.MGH.HARVARD.EDU!Kaplan From: Kaplan@FRODO.MGH.HARVARD.EDU (Josh Kaplan) Newsgroups: bionet.molbio.proteins.7tms_r Subject: endogenous G proteins Date: 9 Oct 1996 05:33:35 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Does anyone know which G alpha subunits are expressed in Xenopus oocytes, and HEK293 cells? I would like to assay the effects of exogenous alpha-o and am shopping for an appropriate cell for expression. COS cells would seem to be ideal due to the overexpression of the transfected construct. Is this the case? Thanks, Josh Joshua M. Kaplan, Ph.D. Department of Molecular Biology Wellman 8 Massachusetts General Hospital 50 Blossom St. Boston, MA 02114 Tel: 617-726-5962 Fax: 617-726-6893 From owner-7tms_r@net.bio.net Tue Oct 08 23:00:00 1996 Path: biosci!BIOCELL.IRMKANT.RM.CNR.IT!7tm From: 7tm@BIOCELL.IRMKANT.RM.CNR.IT ("\\A") Newsgroups: bionet.molbio.proteins.7tms_r Subject: SubstanceK binding Date: 9 Oct 1996 07:42:45 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I'm presently trying to perform receptor binding assays with 125-I-Neurokinin A (Substance K) from Amersham (code n. IM168), using crude membrane preparations from various rat tissues (brain, colon , stomach), but cannot detect any significant binding. I'd really like to know if specific protocols are required for the preparation of membrane samples and/or the handling of the labelled peptide in order to prevent receptor and ligand alteration or degradation. Also, is there a preferential way to sacrify the rats to be used for such membrane preps? Any help greatly welcome! M. Stella Lombardi Inst. Cell Biology CNR I-00137 ROMA Italy ------- From owner-7tms_r@net.bio.net Tue Oct 08 23:00:00 1996 Path: biosci!CHEM.VU.NL!gerhardt From: gerhardt@CHEM.VU.NL (Cindy Gerhardt) Newsgroups: bionet.molbio.proteins.7tms_r Subject: "alpha-adrenergic introns" Date: 9 Oct 1996 14:57:04 -0700 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, I'm interested in the genomic organization of the genes encoding alpha adrenergic receptors. I'm aware of the presence of an intron in the alpha-1c AR gene, but I was wondering if anyone ever found introns in any of the other alpha AR genes? Thanks in advance, Cindy Gerhardt Dept. Biochemistry and Molecular Biology, Vrije Universiteit, Amsterdam, The Netherlands From owner-7tms_r@net.bio.net Wed Oct 09 23:00:00 1996 Path: biosci!ucdavis.edu!cahaskell From: cahaskell@ucdavis.edu (Chris Haskell) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: endogenous G proteins Date: 10 Oct 1996 08:05:25 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 83 Sender: daemon@net.bio.net Distribution: world Message-ID: References: NNTP-Posting-Host: net.bio.net I don't know about COS or HEK cells, but oocytes have the following w/ Gen/EMBL accession numbers: Times =09 Times_New_RomanGSymbola= Times_New_Romano Olate, J X14636 most abundant of GSymbolaTimes_New_Roman isoforms=09 GSymbolaTimes_New_Romans-1a Olate, J X56091 2 isoforms isolated Times Times_New_RomanGSymbolaTimes_New_Romani-1 Olate, J X56089 85% identity to humanTimes Times_New_RomanGSymbolaTimes_New_Romani-3 Olate, J X56090 87% identity to humanTimes Times_New_RomanGSymbolaTimes_New_Romanq Shapira, H Z46958 96% identity to mouseTimes Times_New_RomanGSymbolaTimes_New_Roman11 Shapira, H U10494 92% identity to mouseTimes I believe this list is complete, but if any more are known, please inform. Chris >Does anyone know which G alpha subunits are expressed in Xenopus oocytes, >and HEK293 cells? I would like to assay the effects of exogenous alpha-o >and am shopping for an appropriate cell for expression. COS cells would >seem to be ideal due to the overexpression of the transfected construct. >Is this the case? ----------------------------------------------------------------------- Chris Haskell Dept. Biological Chemistry School of Medicine cahaskell@ucdavis.edu University of California, Davis (916) 752-9070 Davis, CA 95616 =20 ----------------------------------------------------------------------- From owner-7tms_r@net.bio.net Wed Oct 09 23:00:00 1996 Path: biosci!BIOCELL.IRMKANT.RM.CNR.IT!maria From: maria@BIOCELL.IRMKANT.RM.CNR.IT (Maria Stella Lombardi) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subtance K binding Date: 10 Oct 1996 07:14:35 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 16 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I'm presently trying to perform receptor binding assays with 125-I-Neurokinin A (Substance K) from Amersham (code n. IM168), using crude membrane preparations from various rat tissues (brain, colon , stomach), but cannot detect any significant binding. I'd really like to know if specific protocols are required for the preparation of membrane samples and/or the handling of the labelled peptide in order to prevent receptor and ligand alteration or degradation. Also, is there a preferential way to sacrify the rats to be used for such membrane preps? Any help greatly welcome! M. Stella Lombardi Inst. Cell Biology CNR I-00137 ROMA Italy ------- From owner-7tms_r@net.bio.net Wed Oct 09 23:00:00 1996 Path: biosci!BIOCELL.IRMKANT.RM.CNR.IT!maria From: maria@BIOCELL.IRMKANT.RM.CNR.IT (Maria Stella Lombardi) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subtance K binding Date: 10 Oct 1996 02:33:48 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I'm presently trying to perform receptor binding assays with 125-I-Neurokinin A (Substance K) from Amersham (code n. IM168), using crude membrane preparations from various rat tissues (brain, colon , stomach), but cannot detect any significant binding. I'd really like to know if specific protocols are required for the preparation of membrane samples and/or the handling of the labelled peptide in order to prevent receptor and ligand alteration or degradation. Also, is there a preferential way to sacrify the rats to be used for such membrane preps? Any help greatly welcome! M. Stella Lombardi Inst. Cell Biology CNR I-00137 ROMA Italy ------- From owner-7tms_r@net.bio.net Wed Oct 09 23:00:00 1996 Path: biosci!ucdavis.edu!cahaskell From: cahaskell@ucdavis.edu (Chris Haskell) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: endogenous G proteins (readable-sorry) Date: 10 Oct 1996 16:56:06 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 52 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Sorry about that last post. I don't know about COS or HEK cells, but oocytes have the following w/ Gen/EMBL accession numbers: Hetero-trimericTimes G-protein subunitsTimes .Go Olate, J X14636 most abundant of G( isoformsTimes .Gs-1a Olate, J X56091 2 isoforms isolated .Times .Gi-1 Olate, J X56089 85% identity to humanTimes .Gi-3 Olate, J X56090 87% identity to humanTimes .Gq Shapira, H Z46958 96% identity to mouseTimes .G11 Shapira, H U10494 92% identity to mouseTimes Chris ----------------------------------------------------------------------- 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 Fri Oct 11 23:00:00 1996 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 12 Oct 1996 02:00:44 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 239 Sender: daemon@net.bio.net Distribution: world Message-ID: <199610120900.CAA17694@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. 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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 Sat Oct 12 23:00:00 1996 Path: biosci!MUWAYF.UNIMELB.EDU.AU!roger_summers From: roger_summers@MUWAYF.UNIMELB.EDU.AU (Roger Summers) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Sheep beta-adrenoceptor sequences Date: 13 Oct 1996 16:54:21 -0700 Organization: University of Melbourne Lines: 9 Sender: daemon@net.bio.net Distribution: world Message-ID: <3261824F.7964@muwayf.unimelb.edu.au> NNTP-Posting-Host: net.bio.net Does anyone have sequence information for the beta1/2/3 adrenoceptors in the sheep? Many thanks Roger Summers Dept Pharmacology University of Melbourne Parkville Vic 3052 Australia From owner-7tms_r@net.bio.net Sun Oct 13 23:00:00 1996 Path: biosci!agate!howland.erols.net!newsserver.jvnc.net!netnews.sbphrd.com!noddy From: noddy@toytown.org (Noddy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: endogenous G proteins Date: Mon, 14 Oct 1996 10:33:40 +0000 Organization: toytown.org Lines: 14 Distribution: world Message-ID: References: NNTP-Posting-Host: ham992.ha.uk.sbphrd.com Mime-Version: 1.0 Content-Type: text/plain; charset=ISO-8859-1 Content-Transfer-Encoding: 8bit X-Newsreader: Yet Another NewsWatcher 2.2.0b13 In article , Kaplan@FRODO.MGH.HARVARD.EDU (Josh Kaplan) wrote: > Does anyone know which G alpha subunits are expressed in Xenopus oocytes, > and HEK293 cells? I would like to assay the effects of exogenous alpha-o > and am shopping for an appropriate cell for expression. COS cells would > seem to be ideal due to the overexpression of the transfected construct. > Is this the case? > Thanks, > Josh HEK293 cells lack Go. Steve From owner-7tms_r@net.bio.net Sun Oct 13 23:00:00 1996 Path: biosci!WH.BAYER.COM!hyw From: hyw@WH.BAYER.COM (Heeja Yoo-Warren) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Cell lines + B-AR Date: 14 Oct 1996 11:27:26 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: <199610141823.AA27417@miwr03il_e.wh.bayer.com> NNTP-Posting-Host: net.bio.net I am looking for human cell lines which express beta 1 and 2 adrenergic receptors, either singly or both. If you have any information on this, please send me a message at hyw@wh.bayer.com Thank you very much. From owner-7tms_r@net.bio.net Mon Oct 14 23:00:00 1996 Path: biosci!agate!howland.erols.net!www.nntp.primenet.com!nntp.primenet.com!nntp.uio.no!nntp.zit.th-darmstadt.de!fu-berlin.de!informatik.tu-muenchen.de!lrz-muenchen.de!uni-erlangen.de!winx03!wpxx02!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Cell lines + B-AR Date: 15 Oct 1996 08:42:42 GMT Organization: University of Wuerzburg, Germany Lines: 13 Distribution: world Message-ID: <53viq2$u4v@winx03.informatik.uni-wuerzburg.de> References: <199610141823.AA27417@miwr03il_e.wh.bayer.com> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] Heeja Yoo-Warren (hyw@WH.BAYER.COM) wrote: > I am looking for human cell lines which express beta 1 and 2 adrenergic > receptors, either singly or both. If you have any information on > this, please send me a message at hyw@wh.bayer.com The "classical" ones are HEK293 and A431. --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 Mon Oct 14 23:00:00 1996 Path: biosci!SUN.UCHC.EDU!hla From: hla@SUN.UCHC.EDU (Timothy Hla) Newsgroups: bionet.molbio.proteins.7tms_r Subject: subscribe Date: 15 Oct 1996 06:52:17 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 3 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net subscribe From owner-7tms_r@net.bio.net Thu Oct 17 23:00:00 1996 Path: biosci!mail.nrgn.com!rbrodbeck From: rbrodbeck@mail.nrgn.com ("Robbin Brodbeck") Newsgroups: bionet.molbio.proteins.7tms_r Subject: G Protein Content of CHO Date: 18 Oct 1996 13:08:38 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 23 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear GPCR Group: In the interest of better understanding the artificial expression systems we all use to express our various GPCR's, i.e., CHO, Cos, Sf9, etc., I am wondering about G-protein expression. There has been numerous postings in the last month or so dealing with the endogenous receptors expressed in most of the common cell lines used in research, but nothing with respect to the endogenous GPs found! This is obviously a very important issue to us all! I'm wondering if anyone knows of any data on the characterization of GP content for either common research cell lines, or various tissues in the body with particular attention to regions of the brain. I will gladly summarize and post all information I recieve on this subject. Thanks in advance, Robbin Brodbeck Neurogen Corp. From owner-7tms_r@net.bio.net Thu Oct 24 23:00:00 1996 Path: biosci!MAIL.UTEXAS.EDU!wilcoxrich From: wilcoxrich@MAIL.UTEXAS.EDU (Rich E. Wilcox) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Ques. on Koff in intact cells Date: 25 Oct 1996 08:13:26 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 25 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Hello: A colleague and I are trying to model some of our empirical data on desensitization using Monte Carlo methods and we need some kinetic information done at 37 degrees C and using intact cells. Specifically, we would be interested in solid info. on Koff for the following; (a) AR dissociation from G, (b) A dissociation from R, and (c) R dissociation from PKA OR bARK. Alternatively, we could also use the Kon for the respective reactions, since we already have equilibrium (Ka) data using an intact cell prep. Our particular prep for this study was D1A DA receptors stably transfected in rat C6 glioma cells. However, for the simulations Koff or Kon data from any 7tm receptor will probably be sufficient. Thanks, Rich Wilcox wilcoxrich@mail.utexas.edu Richard E. Wilcox, Ph.D. Professor & Doluisio Fellow of Pharmacology College of Pharmacy & Institute for Neuroscience University of Texas Wichita St. , PHR 5.224D Austin, TX 78712-1074 512-471-5199 FAX 512-475-6088 or 512-471-5002 From owner-7tms_r@net.bio.net Sat Oct 26 23:00:00 1996 Path: biosci!DNA.BIO.WARWICK.AC.UK!wp From: wp@DNA.BIO.WARWICK.AC.UK ("Sally Ward") Newsgroups: bionet.molbio.proteins.7tms_r Subject: unsubscribe Date: 27 Oct 1996 07:17:26 -0800 Organization: Biol. Sciences Univ. of Warwick Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Sat Oct 26 23:00:00 1996 Path: biosci!garvan.unsw.edu.au!t.iismaa From: t.iismaa@garvan.unsw.edu.au (Tiina Iismaa) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 27 Oct 1996 18:41:03 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 20 Sender: daemon@net.bio.net Distribution: world Message-ID: <199610280240.NAA17069@gimr.garvan.unsw.EDU.AU> NNTP-Posting-Host: net.bio.net subscribe _______________________________________________________________ Tiina Iismaa, PhD Neurobiology Program Garvan Institute of Medical Research 384 Victoria Street Sydney NSW 2010 Australia Tel: [61-2] 9295 8293 Fax: [61-2] 9295 8281 email: t.iismaa@garvan.unsw.edu.au _______________________________________________________________ From owner-7tms_r@net.bio.net Tue Oct 29 22:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!news-stkh.gsl.net!news.gsl.net!news-peer.gsl.net!news.gsl.net!news.sprintlink.net!news-peer.sprintlink.net!howland.erols.net!newsxfer2.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Ques. on Koff in intact cells Date: Wed, 30 Oct 1996 07:04:09 -0500 Organization: University of Michigan Lines: 66 Message-ID: <32774439.5A44@umich.edu> References: Reply-To: RNeubig@umich.edu NNTP-Posting-Host: host-221.subnet-40.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 3.0Gold (Win95; I) CC: rneubig@umich.edu Rich E. Wilcox wrote: > > Hello: A colleague and I are trying to model some of our empirical data on > desensitization using Monte Carlo methods and we need some kinetic > information done at 37 degrees C and using intact cells. Specifically, we > would be interested in solid info. on Koff for the following; (a) AR > dissociation from G, (b) A dissociation from R, and (c) R dissociation from > PKA OR bARK. Alternatively, we could also use the Kon for the respective > reactions, since we already have equilibrium (Ka) data using an intact cell > prep. Our particular prep for this study was D1A DA receptors stably > transfected in rat C6 glioma cells. However, for the simulations Koff or > Kon data from any 7tm receptor will probably be sufficient. Rich, There is surprisingly little good kinetic data on G protein coupled systems (other than rhodopsin-transducin). At the risk of some serious self-promotion, I can point out our older work on alpha2 adrenergic receptors. There is also elegant work from Larry Sklar on formylpeptide receptors. Jennifer Linderman at Michigan has also been doing kinetic studies with alpha1 adrenergic receptors and she has a good book which summarizes signalling mechanisms with emphasis on kinetics but I don't have the exact citation (authors are Lauffenberger and Linderman). I think one of the big problems in this field is that getting good radiolabelled agonists has been difficult (which was why I chose alpha2 AR as my model system in the first place). We are now trying to use spectroscopic probes of RG interactions to begin to address some of the exact questions you are asking. If you find any other info, I'd be interested to get a summary. Rick Posner, R. G., S. P. Fay, M. D. Domalewski, and L. A. Sklar. 1994. Continuous spectrofluorometric analysis of formyl peptide receptor ternary complex interactions. Mol. Pharmacol. 45:65-73. Neubig, R. R. and L. A. Sklar. 1993. Subsecond modulation of formyl-peptide-linked G proteins by GTPgammaS in permeablized neutrophils. Mol. Pharmacol 43:734-740. Fay, S. P., R. G. Posner, W. N. Swann, and L. A. Sklar. 1991. Real-time analysis of the assembly of ligand, receptor, and G protein by quantitative fluorescence flow cytometry. Biochemistry 30:5066-5075. Thomsen, W. J. and R. R. Neubig. 1989. Rapid kinetics of a2-adrenergic inhibition of adenylate cyclase. Evidence for a distal rate limiting step. Biochemistry 28:8778-8786. Gantzos, R. D. and R. R. Neubig. 1988. Temperature effects on a2-adrenergic receptor-Gi interactions. Biochem. Pharmacol. 37:2815-2821. Neubig, R. R., R. D. Gantzos, and W. J. Thomsen. 1988. Mechanism of agonist and antagonist binding to a2 adrenergic receptors: evidence for a precoupled receptor-guanine nucleotide protein complex. Biochemistry 27:2374-2384. Thomsen, W. J., J. A. Jacquez, and R. R. Neubig. 1988. Inhibition of adenylate cyclase is mediated by the high affinity conformation of the a2-adrenergic receptor. Mol. Pharmacol 34:814-822. _________________________________________________________ Rick Neubig RNeubig@umich.edu Department of Pharmacology University of Michigan Phone (313) 763-3650 FAX (313) 763-4450 http://www-personal.umich.edu/~rneubig/ From owner-7tms_r@net.bio.net Wed Oct 30 22:00:00 1996 Path: biosci!daresbury!nntp-trd.UNINETT.no!news-stkh.gsl.net!news.gsl.net!news-peer.gsl.net!news.gsl.net!howland.erols.net!surfnet.nl!swidir.switch.ch!serra.unipi.it!newsserver.cilea.it!oracle.csi.unimi.it!usenet From: gio@imiucca.csi.unimi.it (Stefano Giovannardi) Newsgroups: bionet.molbio.proteins.7tms_r Subject: test, do not read Date: Thu, 31 Oct 1996 12:42:23 GMT Organization: Universita' degli Studi di Milano Lines: 9 Message-ID: <55a6k1$4rb@oracle.csi.unimi.it> NNTP-Posting-Host: 159.149.32.4 X-Newsreader: Forte Free Agent 1.0.82 Stefano Giovannardi Universita' degli Studi di Milano III Facolta' di Scienze M.F.N. Laboratorio di Fisiologia cellulare e molecolare Via Ravasi 2 21100 Varese, Italy tel: +39-332-250220 fax: +39-332-281308 e-mail: gio@imiucca.csi.unimi.it From owner-7tms_r@net.bio.net Wed Oct 30 22:00:00 1996 Path: biosci!bcm.tmc.edu!cs.utexas.edu!howland.erols.net!newsfeed.internetmci.com!news.webspan.net!ix.netcom.com!netcom.net.uk!sunsite.doc.ic.ac.uk!susx.ac.uk!bafq5 From: bafq5@central.susx.ac.uk (Stephen Perry) Newsgroups: bionet.molbio.proteins.7tms_r Subject: 11-cis-retinal Date: 31 Oct 1996 13:58:53 GMT Organization: University of Sussex Lines: 11 Message-ID: <55abat$ptm@infa.central.susx.ac.uk> NNTP-Posting-Host: solx1.central.susx.ac.uk X-Newsreader: TIN [version 1.2 PL2] Hello, I am trying to acquire a source of high purity 11-cis-retinal for my research. Does anyone know where I can get it from? No companies appear to make it, so a personal contact may be necessary. Thanks in advance Ali Brown From owner-7tms_r@net.bio.net Thu Oct 31 22:00:00 1996 Path: biosci!crtqmai.crtq.com!ricej From: ricej@crtqmai.crtq.com Newsgroups: bionet.molbio.proteins.7tms_r Subject: Unsubsribe Date: 1 Nov 1996 09:33:59 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 9 Sender: daemon@net.bio.net Distribution: world Message-ID: <9610018468.AA846872700@crtqmai.crtq.com> NNTP-Posting-Host: net.bio.net Could someone send me the address to unsubscribe. I have a user that left our company without unsubscribing or changing his email address. I would like to have him removed from your list so that your messages are no longer rejected by our system. Jim Rice Cortech, Inc. jrice@crtq.com