From owner-7tms_r@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!EMBL-HEIDELBERG.DE!Gert.Vriend From: Gert.Vriend@EMBL-HEIDELBERG.DE Newsgroups: bionet.molbio.proteins.7tms_r Subject: GPCRDB server Date: 6 Dec 1995 05:45:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HYHOA01IZI8Y4Y1Z@EMBL-Heidelberg.DE> NNTP-Posting-Host: net.bio.net The GPCRDB server has been updated. In Mosaic or Netscape, use the URL http://swift.embl-heidelberg.de/7tm/ In case something went wrong, all old data is still present under the URL: http://swift.embl-heidelberg.de/7tm1/ Have fun, and please report problems, errors, or suggestions for improvements. Gert. From owner-7tms_r@net.bio.net Tue Dec 05 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!uwm.edu!msunews!netnews.upenn.edu!cronkite.ocis.temple.edu!astro.ocis.temple.edu!driska From: driska@astro.ocis.temple.edu (Stephen P. Driska PhD) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Aluminum fluoride: can it cross an intact plasma membrane? Date: 6 Dec 1995 23:17:02 GMT Organization: Temple University, Academic Computer Services Lines: 19 Message-ID: <4a589e$dva@cronkite.ocis.temple.edu> NNTP-Posting-Host: astro.ocis.temple.edu X-Newsreader: TIN [version 1.2 PL2] The other day I heard someone assert that (AlF4)- was cell-permeable and that it activated G-protein pathways by binding to G-protein alpha subunits that were binding GDP. The story was that because AlF4- was similar in appearance to the phosphate ion and it somehow mimicked the G-protein alpha subunit with GTP bound. My question is how would AlF4- enter intact cells ? It seems that it would be both bulky and negatively charged, and therefore unlikely to enter the cell readily. Does it utilize a phosphate transport system or just use channels or leaks enter ? Just curious. Thanks for any ideas or references. Steve -- Steve Driska, Physiology Department, Temple University Medical School Philadelphia, PA 19140 USA (215) 707-3283 driska@astro.ocis.temple.edu If I could think of something witty and clever to say, it would go right here ----->> ......................... From owner-7tms_r@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!fdn.fr!jussieu.fr!univ-lille1.fr!cresimm-pc3.univ-lille1.fr!vergoten From: vergoten@pop.univ-lille1.fr (Gerard Vergoten) Newsgroups: bionet.molbio.proteins.7tms_r Subject: NATO-ASI on Biomolecular structure and dynamics"NATO Advanced Study Institute" Date: Thu, 7 Dec 1995 10:02:25 Organization: CRESIMM - C8 - USTL - Villeneuve d'Ascq Lines: 16 Distribution: world Message-ID: NNTP-Posting-Host: cresimm-pc3.univ-lille1.fr Keywords: molecular modeling, computer simulations X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] "BIOMOLECULAR STRUCTURE AND DYNAMICS: RECENT EXPERIMENTAL AND THEORETICAL ADVANCES" ASI LOUTRAKI, Greece May 27-June 6,1996 Contact: Professor G. VERGOTEN Address: Universiti des Sciences et Technologies de Lille CRESIMM ( U 279 INSERM ) UFR de Chimie Bbt C8 - 1er itage 59655 VILLENEUVE D'ASCQ FRANCE FAX : (33) 20 33 72 79 E mail : vergoten@pop.univ-lille1.fr Designated Publisher: KLUWER From owner-7tms_r@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!YALE.EDU!virginia.boundy From: virginia.boundy@YALE.EDU ("Virginia Boundy") Newsgroups: bionet.molbio.proteins.7tms_r Subject: FWD>Aluminum fluoride- can Date: 7 Dec 1995 08:22:17 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 26 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Mail*Link(r) SMTP FWD>Aluminum fluoride: can it cross an intact plasma... On 12/7/95, Stephen P. Driska PhD asked about (AlF4)- mimicking the phosphate ion... Here are two references: 1. Bigay J, Deterre P, Pfister C, Chabre M. Fluoride complexes of aluminum or beryllium act on G-proteins as reversibly bound analogues of the g phosphate of GTP. EMBO J 1987; 6:2907-2913. 2. Bigay J, Deterre P, Pfister C, Chabre M. Fluoroaluminates activate transducin-GDP by mimicking the g-phosphate of GTP in its binding site. FEBS Lett 1985; 191:181-185. Hope this is useful. Virginia --- Virginia A. Boundy, Ph.D Molecular Psychiatry - Dept Psychiatry Yale University virginia.boundy@yale.edu From owner-7tms_r@net.bio.net Wed Dec 06 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!elroy.jpl.nasa.gov!swrinde!newsfeed.internetmci.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: FWD>Aluminum fluoride- can Date: Thu, 07 Dec 1995 22:08:39 -0500 Organization: University of Michigan Lines: 29 Message-ID: <30C7AC37.5D2A@umich.edu> References: NNTP-Posting-Host: pm037-06.dialip.mich.net Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b3 (Win95; I) Virginia Boundy wrote: > > On 12/7/95, Stephen P. Driska PhD asked about (AlF4)- mimicking the phosphate > ion... > 2. Bigay J, Deterre P, Pfister C, Chabre M. Fluoroaluminates activate > transducin-GDP by mimicking the g-phosphate of GTP in its binding site. FEBS > Lett 1985; 191:181-185. > There are now crystal structures of both G-alpha-i1 and transducin complexed with GDP and AlF4- Here's a header from the Protein Data Bank file for G alpha-i1 HEADER SIGNAL TRANSDUCTION PROTEIN 11-NOV-94 1GFI 1GFI COMPND GUANINE NUCLEOTIDE-BINDING PROTEIN G(I), ALPHA-1 SUBUNIT 1GFI COMPND 2 (G(I)-ALPHA-1) (ACTIVE FORM) COMPLEXED WITH GDP-ALF4 1GFI SOURCE ORGANISM: RAT (RATTUS RATTUS); EXPRESSION SYSTEM: 1GFI SOURCE 2 ESCHERICHIA COLI; PLASMID: PQE6 VECTOR 1GFI AUTHOR D.E.COLEMAN,A.M.BERGHUIS,S.R.SPRANG 1GFI REVDAT 2 15-MAY-95 1GFIA 1 JRNL REVDAT 1 31-MAR-95 1GFI 0 1GFI JRNL AUTH D.E.COLEMAN,A.M.BERGHUIS,E.LEE,M.E.LINDER, 1GFI JRNL AUTH 2 A.G.GILMAN,S.R.SPRANG 1GFI JRNL TITL STRUCTURES OF ACTIVE CONFORMATIONS OF G=I ALPHA 1= 1GFI JRNL TITL 2 AND THE MECHANISM OF GTP HYDROLYSIS JRNL REF SCIENCE V. 265 1405 1994 1GFI JRNL REFN ASTM SCIEAS US ISSN 0036-8075 0038 1GFI Rick Neubig From owner-7tms_r@net.bio.net Thu Dec 07 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!chi-news.cic.net!nntp.coast.net!swidir.switch.ch!scsing.switch.ch!news.belwue.de!fu-berlin.de!cs.tu-berlin.de!uni-erlangen.de!winx03!wpxx02.toxi.uni-wuerzburg.de!not-for-mail From: krasel@wpxx02.toxi.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Sucrose monolaureate Date: 6 Dec 1995 22:12:41 GMT Organization: Dept. of Pharmacology, U Wuerzburg Lines: 57 Message-ID: <4a54gp$ncj@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: wpxx02.toxi.uni-wuerzburg.de X-Newsreader: TIN [UNIX 1.3 950824BETA PL0] I have to apologize in advance for cluttering up this newsgroup with a private reply, but I couldn't send Steve a private reply (Microsoft Mail bounced it); besides, it might be interesting for others as well. Sorry. Steve asked in private email (> > is something I posted here a few weeks (?) ago): > In article <48tdta$acl@winx03.informatik.uni-wuerzburg.de>, you wrote: > > > > We use laurylsucrose (which is AFAIK the same) in the purification of > > recombinant beta2-adrenergic receptors expressed in Sf9 cells. The > > cell membranes are solubilized in laurylsucrose which is later > > exchanged with dodecylmaltoside. > > > Do you have any more details on these studies in publications? What was > the criteria you used in your choice of detergents (maintainence of > binding? other?) Why the exchange of sucrose monolaureate for > dodecylmaltoside? The receptor is purified via an antagonist affinity column. For some odd reason, the receptor solubilized in DM doesn't bind to the column very well, so it is solubilized in LS and thus loaded on the column. If I recall correctly, we wash then with DM and elute similarly. I don't know if this ever has been published. You *might* be able to find it in Kleymann et al, EJB 213 (1993), 797-804 (not by us). > We recently put a "His-Tag" on the C-terminal of the thrombin receptor and > wish to isolate a photolabeled receptor. I'm wondering if the detergent > micells are covering up the "His-Tag" and I'm wondering about other choices > of detergents. If you don't have it already, you might like to have a look at the following paper: @article{janssen:95, author = {Jacques J. M. Janssen and Petra H. M. Bovee-Geurts and Maarten Merkx and Willem J. DeGrip}, title = {Histidine Tagging Both Allows Convenient Single-step Purification of Bovine Rhodopsin and Exerts Ionic Strength-dependent Effects on Its Photochemistry.}, journal = JBC, volume = 270, number = 19, pages = {11222--11229}, year = 1995 } Hope that helps, --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 Thu Dec 07 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!agate!howland.reston.ans.net!nntp.coast.net!swidir.switch.ch!swsbe6.switch.ch!news.vub.ac.be!bioftp.unibas.ch!daresbury!not-for-mail From: mcroos@imb-jena.de (Martin Roos) Newsgroups: bionet.molbio.proteins.7tms_r Subject: AlF4- : any lab-experiences? Date: 8 Dec 1995 16:10:57 -0000 Lines: 41 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <4a9o2h$mrc@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk To enter into the discussion about the use of AlF4- in receptor research, started by Stephen P. Driska, PhD, 12/7/95: Concerning the beta-adrenergic receptor, it is already textbook knowledge, that an Gsalpha in the "on" form reduces the affinity of the receptor for its hormon ligand. I wonder: 1. if this principle can be applied as well for other G-protein coupled receptors. 2. if this principle could be applied for isolating receptor, ex vivo, with low amount of endogenous ligand bound to the receptor. I am working with endothelin-receptors, where once bound endothelin is nearly undissociable. This fact impares the yield of affinity-purified receptor, using endothelin as ligand. Instead of expensive GTP I have been thinking about AlF4- to be the molecule keeping the G-Protein "on". Any comments, suggestions? Martin ________________________________________________ Martin Roos, PhD-student Institut fuer Molekulare Biotechnologie e.V Proteinlabor Beutenbergstr. 11 D-07745 Jena Germany e-mail: mcroos@imb-jena.de fax: ++49-3641-656225 tel: ++49-3641-656263 __________________________________________________________ From owner-7tms_r@net.bio.net Fri Dec 08 22:00:00 1995 Path: biosci!ihnp4.ucsd.edu!news.service.uci.edu!usenet From: Rob Steele Newsgroups: bionet.molbio.proteins.7tms_r Subject: (no subject) Date: 8 Dec 1995 23:31:08 GMT Organization: University of California, Irvine Lines: 11 Message-ID: <4aahrs$haj@news.service.uci.edu> NNTP-Posting-Host: xenopus.biochem.uci.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Macintosh; I; 68K) X-URL: news:bionet.molbio.proteins.7tms_r We are studying development of the cnidarian Hydra. In the process of cloning homeobox genes from this animal we have recently identified a homologue of PAX-6, a paired-type gene involved in eye development in flies and mammals. Since cnidarians have primitive eyes, we have become interested in the possibility of identifying an opsin gene in Hydra. Is anyone aware of a successful PCR approach for specifically cloning members of the opsin subfamily of 7TM receptors? If so, could you send me the reference (if it has been published) or details on the method if it hasn't yet been published. Thanks for your help. From owner-7tms_r@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!btnet!demon!cityscape.co.uk!usenet From: ae55@cityscape.co.uk (John Bates) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Protein-Bound NSAID problem Date: 11 Dec 1995 13:48:01 GMT Organization: IP-GOLD User Lines: 12 Message-ID: <4ahcqh$18b@news.cityscape.co.uk> NNTP-Posting-Host: cisae55.demon.co.uk X-Newsreader: WinVN 0.92.6+ Several of the NSAID drugs are very hydrophobic and readily bind to hydrophobic sites on certain plasma proteins, particularly albumin. This is creating a problem for a flurbiprofen/ibuprofen plasma extraction technique I am using. Any suggestion on how these drugs can be released from these sites. I don't want to use an organic such as methanol. I have used the conventional liquid-liquid and solid phase techniques successfully and in fact they are very simple. The specialised technique I am using however is somewhat different and requires that I "release" the two NSAIDs before extraction. John From owner-7tms_r@net.bio.net Sun Dec 10 22:00:00 1995 Path: biosci!FARMR1.MED.UTH.TMC.EDU!aschonb From: aschonb@FARMR1.MED.UTH.TMC.EDU (A. Schonbrunn) Newsgroups: bionet.molbio.proteins.7tms_r Subject: PTX sensitive G proteins Date: 11 Dec 1995 07:34:30 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 14 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net I would appreciate recommendations for review articles summarizing differences/similarities in the biochemical and signalling properties of the pertussis toxin sensitive G proteins. Thanks in advance. ************************************************************************** Agi Schonbrunn, Ph.D. e-mail: aschonb@farmr1.med.uth.tmc.edu Dept. of Pharmacology, Univ. of Texas Medical School P.O.Box 20708, Houston,TX 77225 From owner-7tms_r@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!internet!biosci!not-for-mail From: biohelp (BIOSCI Administrator) Newsgroups: bionet.molbio.proteins.7tms_r Subject: IMPORTANT: BIOSCI miniFAQ Date: 12 Dec 1995 02:01:18 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 202 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512121000.CAA07523@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 08-DEC-95) This is a new "miniFAQ" designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. Contents: -------- 1) What to do about "spams," i.e., junk mail, ads, etc. 2) Examples of subscribing and unsubscribing to the mailing lists. 3) How to access BIOSCI/bionet newsgroup archives. 4) The BIOSCI user address and research interest directory. 1) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups) and mailing lists. The same postings are distributed on both 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. 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E-mail users can search the BIOSCI archives by using our waismail e-mail server. For instructions send the message help to waismail@net.bio.net. Leave the Subject: line blank (anything entered on the Subject: line is ignored). 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-7tms_r@net.bio.net Mon Dec 11 22:00:00 1995 Path: biosci!sci.himeji-tech.ac.jp!yagisawa From: yagisawa@sci.himeji-tech.ac.jp (YAGISAWA, Hitoshi) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Subscription Date: 12 Dec 1995 03:12:37 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 13 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512121116.UAA27931@perm.sci.himeji-tech.ac.jp> NNTP-Posting-Host: net.bio.net Subscribe Hitoshi YAGISAWA -------------------------------- Hitoshi YAGISAWA, Ph.D Laboratory of Biological Signalling Department of Life Science Himeji Institute of Technology Harima Science Garden City Hyogo, 678-12 JAPAN Tel: +81-7915-8-0204 Fax: +81-7915-8-0197 ------------------------------- From owner-7tms_r@net.bio.net Tue Dec 12 22:00:00 1995 Path: biosci!PO.CWRU.EDU!pre From: pre@PO.CWRU.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: AlF4- : any lab-experiences? Date: 13 Dec 1995 07:00:33 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512131458.JAA02805@slc5.INS.CWRU.Edu> NNTP-Posting-Host: net.bio.net > To enter into the discussion about the use of AlF4- in receptor research, > started by Stephen P. Driska, PhD, 12/7/95: > > I am working with endothelin-receptors, where once bound endothelin is > nearly undissociable. This fact impares the yield of affinity-purified > receptor, using endothelin as ligand. > Instead of expensive GTP I have been thinking about AlF4- to be the > molecule keeping the G-Protein "on". > Endothelin binding is not entirely G-protein dependent. Have you tried eluting with an alkaline buffer? Endothelin binding is highly pH-sensitive. Also, DTT will denature endothelin owing to the critical disulfide bridges without harming the receptor. -- Paul Ernsberger, PhD, Associate Prof. of Medicine, Pharmacology & Neuroscience Case Western Reserve University, Cleveland, OH 44106-4982 FAX: (216)368-4752 Address all e-mail to: pre@po.cwru.edu *+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+*+* From owner-7tms_r@net.bio.net Thu Dec 14 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!howland.reston.ans.net!newsserver.jvnc.net!synapse.bms.com!pawlowski.bms.com!user From: pawlowski@bms.com (John Pawlowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Largest GPCR agonists? Date: 15 Dec 1995 21:04:24 GMT Organization: BMS Lines: 13 Message-ID: NNTP-Posting-Host: pawlowski.bms.com Having recently attended a conference on GPCRs which was heavily devoted to ligand recognition, structure/function relationships and modeling, I was wondering about the differences in ligands of GPCRs versus ligands of tyrosine kinase receptors. In particular, the peptide ligands of GPCRs are quite small, whereas TKR agonists are generally >10 kDa. I was wondering if anyone could tell me what is the largest GPCR ligand (thrombin doesn't count), and what are the smallest activators of TKRs? Thanks much. John -- John Pawlowski pawlowski@bms.com From owner-7tms_r@net.bio.net Thu Dec 14 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in1.uu.net!nih-csl!helix.nih.gov!dewolf From: "A.M. van Rhee" Subject: GPCR database Content-Type: TEXT/PLAIN; charset=US-ASCII Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: helix.nih.gov Organization: National Institutes of Health Mime-Version: 1.0 Date: Fri, 15 Dec 1995 17:27:58 GMT Lines: 26 Oops, sorry about that. I apparently forgot that some browsers are very sensitive to the exact syntax of the URL. The URL should be: http://mgddk1.niddk.nih.gov:8000/GPCR.html and don't use a trailing / (that leads to the source text and won't have active links). Sorry again, Michael. Dr. A.M. van Rhee National Institutes of Health National Institute of Diabetes, Digestive and Kidney Diseases Laboratory of Bioorganic Chemistry - Molecular Recognition Section Bldg. 8A Rm. B1A17 Bethesda, Maryland 20892-0810 tel: (301) 435-1936 // fax: (301) 402-4182 // dewolf@helix.nih.gov URL: http://mgddk1.niddk.nih.gov:8000/ From owner-7tms_r@net.bio.net Thu Dec 14 22:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!in2.uu.net!nih-csl!helix.nih.gov!dewolf From: "A.M. van Rhee" Subject: GPCR Mutant Database Content-Type: TEXT/PLAIN; charset=US-ASCII Message-ID: Sender: postman@alw.nih.gov (AMDS Postmaster) Nntp-Posting-Host: helix.nih.gov Organization: National Institutes of Health Mime-Version: 1.0 Date: Fri, 15 Dec 1995 13:29:55 GMT Lines: 41 Hello all. A short while ago I posted something about a mutant GPCR database that we're working on here at NIH. Because of high demand, we've speeded up the process of conversion to HTML and we now have a part of our database on-line. The way it's set-up is not ideal (not even for our own standards), but it will allow you to find out more about mutants in the transmembrane domains of family A/rhodopsin related GPCRs. Since it's all very experimental, we'd really appreciate comments and contributions from the GPCR community at large. We've been talking to Frank Kolakowski (GCRdB) and Gert Vriend (EMBL) to set up a common site/framework for this kind of database, but I'd really like to get some input from other people. Besides, I think a common project would take at least half a year to get started, and I don't want to keep everyone in suspense. You can access the database through URL: http://mgddk1.niddk.nih.gov:8000/GPCR.html/ Have fun with it, and hope to hear from you soon. Michael. Dr. A.M. van Rhee National Institutes of Health National Institute of Diabetes, Digestive and Kidney Diseases Laboratory of Bioorganic Chemistry - Molecular Recognition Section Bldg. 8A Rm. B1A17 Bethesda, Maryland 20892-0810 tel: (301) 435-1936 // fax: (301) 402-4182 // dewolf@helix.nih.gov URL: http://mgddk1.niddk.nih.gov:8000/ From owner-7tms_r@net.bio.net Fri Dec 15 22:00:00 1995 Path: biosci!agate!howland.reston.ans.net!swrinde!cssun.mathcs.emory.edu!emory!news.cc.emory.edu!usenet From: medtjm@bimcore.emory.edu (T. J. Murphy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Largest GPCR agonists? Date: 16 Dec 1995 21:42:49 GMT Organization: Biomolecular Computing Resource, Emory University Lines: 15 Distribution: world Message-ID: <4avegp$sgq@moe.cc.emory.edu> References: Reply-To: medtjm@bimcore.emory.edu NNTP-Posting-Host: bimcore.emory.edu In article 1512951736280001@pawlowski.bms.com, pawlowski@bms.com (John Pawlowski) writes: >..... I was wondering >if anyone could tell me what is the largest GPCR ligand (thrombin doesn't >count), and what are the smallest activators of TKRs? Thanks much. > You didn't ask, but my favorite smallest activator of a GPCR is the photon! --- TJ Murphy Asst. Professor Dept of Pharmacology Emory University School of Medicine From owner-7tms_r@net.bio.net Sat Dec 16 22:00:00 1995 Path: biosci!VAX.GRC.NIA.NIH.GOV!chahrzad From: chahrzad@VAX.GRC.NIA.NIH.GOV Newsgroups: bionet.molbio.proteins.7tms_r Subject: Gi-linked receptor Date: 17 Dec 1995 11:42:35 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Distribution: world Message-ID: <0099B016.226A989A.12@vax.grc.nia.nih.gov> NNTP-Posting-Host: net.bio.net Could anyone help me clarify this issue for me? I have a peptide that in. could some tissues lowers cAMP but via a pertussis toxin insensitive manner! How could this be? Please let me know if you have any information on similar situations with other peptides or hormones and please let me know of elase already published data (if available). Thank you! Chahrzad Montroseo Email: Chahrzad@vax.grc.nia.nih.govrhr From owner-7tms_r@net.bio.net Sun Dec 17 22:00:00 1995 Path: biosci!novo.dk!khou From: khou@novo.dk (Khaled M. Houamed) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Largest GPCR agonists? Date: 18 Dec 1995 12:23:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net >Having recently attended a conference on GPCRs which was heavily devoted >to ligand recognition, structure/function relationships and modeling, I >was wondering about the differences in ligands of GPCRs versus ligands of >tyrosine kinase receptors. In particular, the peptide ligands of GPCRs are >quite small, whereas TKR agonists are generally >10 kDa. I was wondering >if anyone could tell me what is the largest GPCR ligand (thrombin doesn't >count), and what are the smallest activators of TKRs? Thanks much. > >John > >-- >John Pawlowski >pawlowski@bms.com The glycoprotein hormones LH, FSH, TSH are > 20 kilo-dalton From owner-7tms_r@net.bio.net Sun Dec 17 22:00:00 1995 Path: biosci!CTS.COM!Kinta From: Kinta@CTS.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: Detection of intracellular cAML levels Date: 18 Dec 1995 14:43:25 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 10 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Dear Netter: Dose anyone know the methods for direct detection of intracelluar cAMP levels? I'm looking for the technique to visualize (color or fluorescence etc.) the intracellular cAMP levels. Any help would be appreciated. S. Kaneta From owner-7tms_r@net.bio.net Sun Dec 17 22:00:00 1995 Path: biosci!PINK.INCM.U-NANCY.FR!campagne From: campagne@PINK.INCM.U-NANCY.FR (Fabien Campagne) Newsgroups: bionet.molbio.proteins.7tms_r Subject: HELP Date: 18 Dec 1995 02:26:52 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 8 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Fabien Campagne -- campagne@pink.incm.u-nancy.fr | Theoretical Chemistry Dept. phone: (033) 83 91 20 00 extension 3236 | University of Nancy, France. From owner-7tms_r@net.bio.net Sun Dec 17 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!usenet.eel.ufl.edu!clas.ufl.edu!usenet.ufl.edu!receptor.pharmacology.ufl.edu!user From: jharrison@qm.server.ufl.edu (Jeffrey K. Harrison) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Largest GPCR agonists? Followup-To: bionet.molbio.proteins.7tms_r Date: Mon, 18 Dec 1995 10:18:15 -0500 Organization: University of Florida Lines: 11 Message-ID: References: NNTP-Posting-Host: receptor.pharmacology.ufl.edu How about chemoattractant cytokines (chemokines)? Not sure if they are the largest, but they are probably near the top of the list. These ligands are 70-80 amino acids in length, and about 8-10 kDa in size. Some of the rodent orthologs are even larger, e.g. murine/rat MCP-1, weighing in around 14 kDa. Jeffrey K. Harrison, Ph.D. Assistant Professor of Pharmacology and Therapeutics University of Florida From owner-7tms_r@net.bio.net Mon Dec 18 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: Detection of intracellular cAML levels Date: Tue, 19 Dec 1995 10:40:36 -0500 Organization: University of Michigan Lines: 36 Message-ID: <30D6DCF4.12C1@umich.edu> References: NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b3 (Win95; I) Kinta@CTS.COM wrote: > > Dear Netter: > > Dose anyone know the methods for direct detection of intracelluar cAMP levels? > I'm looking for the technique to visualize (color or fluorescence etc.) the > intracellular cAMP levels. Tsien and Taylor published a method based on energy transfer between the regulatory and catalytic subunits of PKA. I'm not sure how easy this is but check the following references. (I seem to recall seeing a commercial advertisement for reagents for this method). TI: Imaging of cAMP signals and A-kinase translocation in single living cells. AU: Adams-S; Bacskai-B; Harootunian-AT; Mahaut-Smith-M; Sammak-PJ; Taylor-SS; Tsien-RY SO: Adv-Second-Messenger-Phosphoprotein-Res. 1993; 28: 167-70 TI: Fluorescence ratio imaging of cyclic AMP in single cells. AU: Adams-SR; Harootunian-AT; Buechler-YJ; Taylor-SS; Tsien-RY SO: Nature. 1991 Feb 21; 349(6311): 694-7 Rick > > Any help would be appreciated. > > S. Kaneta -- _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 From owner-7tms_r@net.bio.net Tue Dec 19 22:00:00 1995 Path: biosci!BIO.VU.NL!hind From: hind@BIO.VU.NL (Hind van Tol-Steye) Newsgroups: bionet.molbio.proteins.7tms_r Subject: intracellular cAMP levels Date: 20 Dec 1995 07:39:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 32 Sender: daemon@net.bio.net Distribution: world Message-ID: <9512201537.AA00749@bio.vu.nl> NNTP-Posting-Host: net.bio.net Dose anyone know the methods for direct detection of intracelluar cAMP levels? > I'm looking for the technique to visualize (color or fluorescence etc.) the > intracellular cAMP levels. Tsien and Taylor published a method based on energy transfer between the regulatory and catalytic subunits of PKA. I'm not sure how easy this is but check the following references. (I seem to recall seeing a commercial advertisement for reagents for this method). TI: Imaging of cAMP signals and A-kinase translocation in single living cells. AU: Adams-S; Bacskai-B; Harootunian-AT; Mahaut-Smith-M; Sammak-PJ; Taylor-SS; Tsien-RY SO: Adv-Second-Messenger-Phosphoprotein-Res. 1993; 28: 167-70 TI: Fluorescence ratio imaging of cyclic AMP in single cells. AU: Adams-SR; Harootunian-AT; Buechler-YJ; Taylor-SS; Tsien-RY SO: Nature. 1991 Feb 21; 349(6311): 694-7 Rick > > Any help would be appreciated. > > S. Kaneta The cAMP fluorosensor developped by Tsien and Taylor is commercially available from Atto instruments, 1450 research boulevard rockville MD 20850 301 340 7320 FAX 301 340 9775, email sales@atto.com. Price is $695.00 for 10 microl of 10-15 microM ready for micro-injection. From owner-7tms_r@net.bio.net Tue Dec 19 22:00:00 1995 Path: biosci!bcm.tmc.edu!news.msfc.nasa.gov!newsfeed.internetmci.com!newsxfer2.itd.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: intracellular cAMP levels Date: Wed, 20 Dec 1995 12:51:34 -0500 Organization: University of Michigan Lines: 14 Message-ID: <30D84D26.3CB5@umich.edu> References: <9512201537.AA00749@bio.vu.nl> NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 2.0b3 (Win95; I) Hind van Tol-Steye wrote: > > The cAMP fluorosensor developped by Tsien and Taylor is commercially > available from Atto instruments, 1450 research boulevard rockville MD 20850 > 301 340 7320 FAX 301 340 9775, email sales@atto.com. Price is $695.00 for > 10 microl of 10-15 microM ready for micro-injection. So have you used it and does it work well? Rick _________________________________________________________ Rick Neubig RNeubig@umich.edu University of Michigan Phone (313) 763-3650 http://www.umich.edu/~rneubig FAX (313) 763-4450 From owner-7tms_r@net.bio.net Wed Dec 20 22:00:00 1995 Path: biosci!BIO.VU.NL!hind From: hind@BIO.VU.NL (Hind van Tol-Steye) Newsgroups: bionet.molbio.proteins.7tms_r Subject: cAMP fluorosensor Date: 21 Dec 1995 06:51:05 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 5 Sender: daemon@net.bio.net Distribution: world Message-ID: <9512211449.AA00324@bio.vu.nl> NNTP-Posting-Host: net.bio.net About the cAMP fluorosensor: I have not used it; we have no equipment to measure emmission ratio's. Hind. From owner-7tms_r@net.bio.net Tue Dec 26 22:00:00 1995 Path: biosci!agate!overload.lbl.gov!news.kreonet.re.kr!news.dacom.co.kr!news.uoregon.edu!news.orst.edu!Schimerlik2-2131-E.ALS.ORST.EDU!bulsecod From: bulsecod@labtalk.als.orst.edu (Lab) Newsgroups: bionet.molbio.proteins.7tms_r Subject: global curve fitting Date: Wed, 27 Dec 1995 10:05:47 Organization: University Computing Services - Oregon State University Lines: 16 Message-ID: NNTP-Posting-Host: schimerlik2-2131-e.als.orst.edu X-Newsreader: Trumpet for Windows [Version 1.0 Rev A] Has anyone used the latest version of Origin (4.0) for non-linear curve fitting? It has 'global' fitting built in (in the sense that you can set it to share parameters across multiple data sets). I am trying to implement this global fitting using different equations, and have not had much success. Anyone out there been able to do this? Also, have you been able to implement 'global fitting' in any other packages besides Scientist and SigmaPlot (and Origin)? Thanks... Dylan bulsecod@labtalk.als.orst.edu From owner-7tms_r@net.bio.net Wed Dec 27 22:00:00 1995 Path: biosci!LYNX.DAC.NEU.EDU!pwick From: pwick@LYNX.DAC.NEU.EDU (peter f wick) Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 28 Dec 1995 13:26:10 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 1 Sender: daemon@net.bio.net Distribution: world Message-ID: <199512282124.QAA28523@lynx.dac.neu.edu> NNTP-Posting-Host: net.bio.net subscribe pwick@lynx.neu.edu 7TMS_r@net.bio.net