From owner-7tms_r@net.bio.net Tue Jun 02 23:00:00 1998 Path: biosci!biosci!not-for-mail From: edvard@fagmed.uit.no (Oyvind Edvardsen) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Mutant database tinyGRAP release 6.0 Date: 3 Jun 1998 05:24:11 -0700 Organization: Univ. of Tromso, IMB Lines: 76 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6l3f9b$70v@net.bio.net> Reply-To: edvard@fagmed.uit.no NNTP-Posting-Host: net.bio.net We are pleased to announce the availability of release 6.0 of tinyGRAP. After a long time we have finally managed to create an update of the database. tinyGRAP release 6.0 is the first real collaborative release between University of Tromso, Leiden Amsterdam Center for Drug Research (LACDR) and the BIOcomputing group at EMBL, Heidelberg. Changes: * Added ~1000 mutants, most notably chimers. * New data file format. * Changed query form to be able to combine receptor information with bibliographic information. Only mutants are returned. * Faster query engine. * Almost every bit of the internal organization of the database has been changed. o Margot Beukers at LACDR has done most of the tedious job of entering data. She has also constructed the data file format and done some serious database 'test-driving'. o Florence Horn at BIOcomputing, EMBL, Heidelberg has contributed to da= ta entry. o Gert Vriend at BIOcomputing, EMBL, Heidelberg has contributed by arra= nging yet another fruitful GPCR meeting (Heidelberg, March this year), by his inter= est in GPCRs and by taking part in and creating interesting discussions. o Oyvind Edvardsen at University of Tromso has created programs to check Margot's data entry and taken care of all the other software and database maintainance stuff. o Margot and Oyvind together are responsible for the remaining bits and pieces. The previous version (5.0) will be available for a while. We would like to thank Dr. K. Kristiansen for useful comments and for being a database 'test-driver'. In order to be able to continue to provide computerized mutant data we need YOUR help. If you can't locate YOUR paper(s) in tinyGRAP, at least send us a notification. Even better if YOU would send reprint(s). The=20 postal address is given below. User feedback is always welcome. NOTE: We are currently logging your queries to be able to discover and ha= ndle NOTE: problems efficiently. NOTE: Query logging will be removed later. Access tinyGRAP from the GRAP homepage: http://www-grap.fagmed.uit.no/GRAP/homepage.html Above is the one and only URL that can be guaranteed to be persistent. =D8yvind. - --=20 - -o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-o-= o-o- ___ _ =20 | /| _| |_ _| __ _ _| _ _ |/_| \/ \/ | |\| |_| |_ |_| \/ |_|_ | |_| _\ |_' |\| / _________________________________________________________________________= ____ School of Medicine | Dept. of Pharmacology, IMB | TelePhone: +47 77 64 53 42 University of Troms=F8 | TeleFax: +47 77 64 53 10 MH, Breivika | Email: edvard@fagmed.uit.no N-9037 TROMS=D8, NORWAY | URL: http://atf1.fagmed.uit.no/mgl.= html - -------------------------------------------------------------------------= - ----- From owner-7tms_r@net.bio.net Mon Jun 08 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Ian" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor solubilization: different detergents Date: 9 Jun 1998 20:04:04 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 105 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6lkt34$6hs@net.bio.net> NNTP-Posting-Host: net.bio.net This is a multi-part message in MIME format. - ------=_NextPart_000_0004_01BD9308.BCB5E2A0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Hello, I have crosslinked a radiolabelled ligand to our receptor in cells. The = cells were solubilised in several detergents (4C) and debris pelletd by = low speed centrifugation.=20 The radiocativity was determined in the pellet and supernatant and = expressed as supernatant over total counts. Detergent % Triton 1% 14 >0.3mM NP-40 16 20mM Na Cholate 33 =20 10mM Deoxychol Na 26 10mM CHAPS 20 Questions: 1. How can I increase solubilisation? 2. How is the micellar weight related to solubilisation?:=20 Cholate and Deoxycol had the lowest micellar wt compared to the other = detergents. Any help or information would be greatly appreciated. Thanks in advance. Sincerely, Ian Murray imurra@po-box.mcgill.ca - ------=_NextPart_000_0004_01BD9308.BCB5E2A0 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Hello,
 
I have crosslinked a radiolabelled ligand to our = receptor in=20 cells. The cells were solubilised in several detergents (4C) and debris = pelletd=20 by low speed centrifugation.
 
The radiocativity was determined in the pellet and = supernatant=20 and expressed as supernatant over total counts.
 
Detergent         &= nbsp;           &n= bsp; =20 %
Triton=20 1%            = ;           =20 14
>0.3mM=20 NP-40           &n= bsp;   =20 16
20mM Na Cholate      = 33    =20
10mM Deoxychol = Na       =20 26
10mM=20 CHAPS           &n= bsp;    =20 20
 
Questions:
1. How can I increase solubilisation?
2. How is the micellar weight related to = solubilisation?:=20
  Cholate and Deoxycol had the lowest micellar = wt=20 compared to the other detergents.
 
Any help or information would be greatly = appreciated. Thanks=20 in advance.
Sincerely,
 
Ian Murray
imurra@po-box.mcgill.ca
- ------=_NextPart_000_0004_01BD9308.BCB5E2A0-- From owner-7tms_r@net.bio.net Tue Jun 09 23:00:00 1998 Path: biosci!biosci!not-for-mail From: WalterThomas Newsgroups: bionet.molbio.proteins.7tms_r Subject: arrestin antibodies. Date: 10 Jun 1998 03:36:10 -0700 Organization: Baker Institute, Melb., Australia Lines: 30 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6llniq$isp@net.bio.net> NNTP-Posting-Host: net.bio.net Hi everyone, I'm trying to determine whether arrestin proteins associate with the angiotensin II (AT1A) receptor in an agonist- and phosphorylation-dependent manner. I recently purchased antibodies against beta-arrestin-1 and beta-arrestin-2 which are distributed by Transduction laboratories. Has anyone used these antibodies successfully to detect arrestin proteins? I ran a Western blot of extracts from a variety of cell lines (HEK293, COS-7, CHO-K1, etc) as well as the internal control extracts supplied with the antibodies and got some very weird results which make me question the specificity of the antibodies. I would be interested in hearing from anyone who has had experience (and success) with these antisera. Does anyone know of any other commercial sources of arrestin antibodies? Thanks Wally Walter G. Thomas Walter.Thomas@baker.edu.au Baker Medical Research Institute Commercial Rd. Prahran, Melbourne Victoria 3181 Australia International Phone 61-3-95224333 International Fax 61-3-95211362 From owner-7tms_r@net.bio.net Wed Jun 17 23:00:00 1998 Path: biosci!biosci!not-for-mail From: Athan Kuliopulos Newsgroups: bionet.molbio.proteins.7tms_r Subject: Two Signal Transduction Postdoc Positions-Boston Date: 18 Jun 1998 11:54:24 -0700 Organization: NEMC Lines: 59 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6mbnp0$64a@net.bio.net> Reply-To: "Kuliopoulos@HemOnc"@NEMC.nemc.org NNTP-Posting-Host: net.bio.net Two Postdoctoral Positions Available Molecular Cardiology Research Institute Tufts University School of Medicine-NEMC Boston, MA We have an immediate opening for two Postdoctoral Fellows to work in the field of molecular signaling and peptide-protein recognition. The projects focus on the human thrombin receptor. The thrombin receptor is activated by thrombin cleavage of the receptor exodomain and exposure of an N-terminal tethered ligand that binds to the body of the 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. 1) Macromolecular Structural Studies of the Resting and Activated States of Thrombin Receptor Extracellular Domains. NMR structural studies of the thrombin receptor exodomain in activated and resting forms are currently in progress and a preliminary structure has been generated for the activated exodomain. Future projects include solving the structure of the exodomain complexed with extracellular loops. Insight into the molecular interactions between the exodomain and the body of the receptor should provide leads for the development of novel anti-thrombin receptor agents in collaboration with a pharmaceutical company. The NMR facility is located in the Medical School Biochemistry Department and current instrumentation include a new Bruker 600 MHz and updated 500 MHz magnets along with several SGI workstations. Our lab has close collaborations with NMR spectroscopists who provide additional technical expertise. 2) Thrombin-Cell Surface Protein Interactions; Development of Cell Surface-Specific Anti-Thrombotic Agents. During coagulation, the physiologic concentration of thrombin exceeds that of its thrombin receptor substrate. Therefore, thrombin has a difficult task of discriminating among the various cell surface proteins to find the receptor and cleave it in the millisecond time range. We are interested in exploring this unusual mechanism of substrate-assisted domain cleavage by thrombin and using this information to develop a novel class of cell surface anti-thrombin agents. The laboratory is located within the Molecular Cardiology Research Institute, a modern, state-of-the-art facility with a staff of 40 investigators including technical support. Qualifications for this position are a Ph.D. degree. Candidates with training in NMR who would like to acquire expertise in molecular biology are 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 of Medicine and Biochemistry Molecular Cardiology Research Institute Tufts-NEMC Box 832 750 Washington Street Boston, MA 02111 617-636-8482 617-636-4833 (fax) akuliopu@opal.tufts.edu From owner-7tms_r@net.bio.net Wed Jun 17 23:00:00 1998 Path: biosci!biosci!not-for-mail From: daaka001@receptor-biol.duke.edu (Yehia Daaka) Newsgroups: bionet.molbio.proteins.7tms_r Subject: postdoc positions Date: 18 Jun 1998 11:53:44 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6mbnno$639@net.bio.net> NNTP-Posting-Host: net.bio.net - --============_-1314040312==_ma============ Content-Type: text/plain; charset="us-ascii" Two postdoctoral positions are available immediately to study the regulation and function of G protein-coupled receptors (see Nature 390, 88, 1997; and J. Biol. Chem. 273, 685, 1998). For studies on regulation, the successful candidate will have experience in recombinant DNA technology. For studies on function, the successful candidate will have experience in molecular biology and/or signal transduction of G protein-coupled receptors and/or receptor tyrosine kinases. Send curriculum vitae and names of three references to: Dr. Yehia Daaka, Medical Science Research Bldg. Box 2621, Duke University Medical Center, Durham, NC, 27710. - --============_-1314040312==_ma============ Content-Type: text/enriched; charset="us-ascii" TimesTwo postdoctoral positions are available immediately to study the regulation and function of G protein-coupled receptors (see Nature 390, 88, 1997; and J. Biol. Chem. 273, 685, 1998). For studies on regulation, the successful candidate will have experience in recombinant DNA technology. For studies on function, the successful candidate will have experience in molecular biology and/or signal transduction of G protein-coupled receptors and/or receptor tyrosine kinases. Send curriculum vitae and names of three references to: Dr. Yehia Daaka, Medical Science Research Bldg. Box 2621, Duke University Medical Center, Durham, NC, 27710. - --============_-1314040312==_ma============-- From owner-7tms_r@net.bio.net Wed Jun 17 23:00:00 1998 Path: biosci!biosci!not-for-mail From: Paul Gouldson Newsgroups: bionet.molbio.proteins.7tms_r Subject: Post Doc / Permenant Position Date: 18 Jun 1998 11:51:33 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6mbnjl$5sp@net.bio.net> NNTP-Posting-Host: net.bio.net Dear All, I am looking for a post doc or perm. position in the subject area of GPCRs. I am currently a post doc at Sanofi Recherche, working for Dr David Shire. I have been investigating small ligand binding sites in different GPCRs using various techniques. I have experinece in the following areas, radio ligand binding assays, cell culture, transient expression in mamalian and bacterial cells, gene construction, GFP tagging and fluorecence microscopy, molecular modelling, molecular dynamics simulations. Addiotionally, I have been extending the work of my Ph.D. thesis, Modelling GPCR Dimers: by investigating further dimerisation in GPCRs. I am looking for a position where i can utilise my knowledge of GPCRs, preferably in the UK. Please mail me for a copy of my current CV. many thanks Paul Gouldson PAUL.GOULDSON@sanofi.com From owner-7tms_r@net.bio.net Sun Jun 28 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "PH" Newsgroups: bionet.molbio.proteins.7tms_r Subject: adrenomedullin receptor Date: 29 Jun 1998 08:44:52 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 12 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6n8cpk$64u@net.bio.net> NNTP-Posting-Host: net.bio.net Dear all ! I'm looking for cell-lines expressing the adrenomedullin- and the proadrenomedullin receptor. Can anyone help ?? Philip Hasbak, MD Dept. of Nuclear medicine Gentofte Hospital University of Copenhagen Denmark E-mail: Philip@post1.tele.dk From owner-7tms_r@net.bio.net Sun Jun 28 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "PH" Newsgroups: bionet.molbio.proteins.7tms_r Subject: Cell lines expressing the AM and PAMP receptors Date: 29 Jun 1998 08:46:15 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 46 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6n8cs7$70e@net.bio.net> NNTP-Posting-Host: net.bio.net This is a multi-part message in MIME format. - ------=_NextPart_000_000D_01BD9DEF.79B82DE0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable Dear Sirs! I'm looking for cell-lines expressing the adrenomedullin- and the PAMP = receptors. Can anybody help ? Philip Hasbak, MD Dept. of Nuclear medicine Gentofte Hospital Univesity of Copenhagen Denmark E-mail: Philip@post1.tele.dk - ------=_NextPart_000_000D_01BD9DEF.79B82DE0 Content-Type: text/html; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable
Dear Sirs!

I'm looking for cell-lines expressing the=20 adrenomedullin- and the PAMP receptors.
Can anybody help = ?


Philip=20 Hasbak, MD
Dept. of Nuclear medicine
Gentofte = Hospital
Univesity of=20 Copenhagen
Denmark
E-mail: Philip@post1.tele.dk
  - ------=_NextPart_000_000D_01BD9DEF.79B82DE0-- From owner-7tms_r@net.bio.net Sun Jun 28 23:00:00 1998 Path: biosci!biosci!not-for-mail From: "Pieter van Santen" Newsgroups: bionet.molbio.proteins.7tms_r Subject: The Main Metabolic Pathways Date: 29 Jun 1998 08:48:14 -0700 Organization: World Access Lines: 6 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6n8cvu$7cu@net.bio.net> NNTP-Posting-Host: net.bio.net Dear colleague, If you are interested in The Main Metabolic Pathways on the Internet, visit: http://home.wxs.nl/~pvsanten/mmp/mmp.html Good luck Pieter From owner-7tms_r@net.bio.net Sun Jun 28 23:00:00 1998 Path: biosci!biosci!not-for-mail From: Marc Andelman Newsgroups: bionet.molbio.proteins.7tms_r Subject: Group leader, G protein coupled receptor cloning Date: 29 Jun 1998 08:47:20 -0700 Organization: Biosource Lines: 21 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6n8cu8$77p@net.bio.net> NNTP-Posting-Host: net.bio.net Biosource is the oldest recruitment firm in the biotechnology industry. Our client, a California based company, would like to hire someone to start a group in G protein coupled receptor molecular biology and cloning. They would especially like to find orphan receptors. This company is well established in the field of high throughput screening, so there should also be some nice interaction and support with other groups in the company to uncover ligands. Intersted people please reply to Marc Andelman below; Regards, Marc Andelman Biosource Inc. tel 508 853 8803 fax 508 853 8772 Email drgonfly@ultranet.com From owner-7tms_r@net.bio.net Sun Jun 28 23:00:00 1998 Path: biosci!biosci!not-for-mail From: Aaron Roseberry Newsgroups: bionet.molbio.proteins.7tms_r Subject: HEK293 Library Date: 29 Jun 1998 08:46:52 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 7 Sender: daemon@net.bio.net Approved: lfk@gcrdb.uthscsa.edu Distribution: world Message-ID: <6n8ctc$751@net.bio.net> NNTP-Posting-Host: net.bio.net Does anyone have an HEK293 cDNA library (or even genomic)? I've looked around in the literature and can't seem to find any mention of one. Aaron Roseberry Northwestern University Chicago Il 60611 (312) 503-2737