From owner-7tms_r@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!usenet.ins.cwru.edu!usenet From: Bryan Roth Newsgroups: bionet.molbio.proteins.7tms_r Subject: rhodopsin coordinates Date: 3 May 1995 18:55:49 GMT Organization: Case Western Reserve University, Cleveland, Ohio (USA) Lines: 8 Message-ID: <3o8jjl$1ko@usenet.INS.CWRU.Edu> NNTP-Posting-Host: meds20785.meds.cwru.edu Some time ago, Baldwin published a model of rhodopsin based on her 2-D crystals. Does anyone know how of if the coordinates of this model may be obtained. Bryan Roth Department of Biochem CWRU roth@biocserver.cwru.edu From owner-7tms_r@net.bio.net Tue May 02 23:00:00 1995 Path: biosci!novo.dk!bsw From: bsw@novo.dk (Birgitte Schjellerup Wulff) Newsgroups: bionet.molbio.proteins.7tms_r Subject: HEK 293 cells and receptors Date: 3 May 1995 23:33:30 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 17 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505040635.AA11220@nngate.novo.dk> NNTP-Posting-Host: net.bio.net Hi Have anyone experience with endogenous 7TM receptors in HEK 293 cells. I'm especially interested in knowing if receptors from the secretin/VIP receptor family is endogenous expressed in the cells. Best regards Birgitte Wulff Birgitte Wulff >bsw@novo.dk< Dept. of Molecular Pharmacology 1C.S36, Pharmaceutical Biotechnology Novo-Nordisk 2880 Bagsvaerd, Denmark tlf: +45 44 42 34 21 fax: +45 44 98 50 07 From owner-7tms_r@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!SYNAPSE.BMS.COM!boundy From: boundy@SYNAPSE.BMS.COM (Virginia A. Boundy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: HEK 293 cells and receptors Date: 4 May 1995 11:02:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505041801.AA01703@synapse> NNTP-Posting-Host: net.bio.net endogenous 7TM receptors in HEK 293 cells.. Sorry, I missed the word endogenous before... D1 dopamine receptors are endogenously expressed at realtively low densities in HEK 293 cells. They do couple to stimulation of adenylyl cyclase in our hands -*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*- Virginia Boundy Dept of Pharmacology University of Pennsylvania Philadelphia, PA 19104 -- CNS Exploratory Bristol-Myers Squibb Pharmaceutical Research Institute Wallingford, CT 06492 (203) 284-6927 From owner-7tms_r@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!SYNAPSE.BMS.COM!boundy From: boundy@SYNAPSE.BMS.COM (Virginia A. Boundy) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: HEK 293 cells and receptors Date: 4 May 1995 09:31:27 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 36 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505041630.AA20650@synapse> NNTP-Posting-Host: net.bio.net In response to bsw@novo.dk (Birgitte Schjellerup Wulff) > >Have anyone experience with endogenous 7TM receptors in HEK 293 cells. I'm >especially interested in knowing if receptors from the secretin/VIP >receptor family is endogenous expressed in the cells. > We (Perry Molinoff's lab) have expressed the human D2 dopamine receptors (both isoforms), the rat D3 dopamine receptors and the beta 2 adrenergic receptor in HEK 293 cells. All the receptors expressed at reasonable densities, bound ligands with appropriate affinities, and shown the expected rank order of potency for agonists and antagonists. The D2 dopamine receptors and the beta 2 adrenergic receptor also coupled to adenylyl cyclase in these cells. We studied the regulation of dopamine receptors in these cells. References: Filtz et al Mol Pharm 44:371-379, 1993 Filtz et al JPET 271:1574-1582, 1994 Burris et al JPET 268:935-942, 1994 Burris et al Neuropsychopharm., in press -*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*- Virginia Boundy Dept of Pharmacology University of Pennsylvania Philadelphia, PA 19104 -- CNS Exploratory Bristol-Myers Squibb Pharmaceutical Research Institute Wallingford, CT 06492 (203) 284-6927 From owner-7tms_r@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!RECEPTOR.MGH.HARVARD.EDU!lfk From: lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: JBC 5_12 TOC for 7tms_R Date: 4 May 1995 05:22:31 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 45 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505041215.AA01374@receptor> NNTP-Posting-Host: net.bio.net AU Leibovitz-D. Koch-Y. Fridkin-M. Pitzer-F. Zwickl-P. Dantes-A. Baumeister-W. Amsterdam-A. TI Archaebacterial and eukaryotic proteasomes prefer different sites in cleaving gonadotropin-releasing hormone SO J-Biol-Chem. 1995 May 12. 270(19). p 11029. AU Molino-M. Blanchard-N. Belmonte-E. Tarver-A-P. Abrams-C. Hoxie-J-A. Cerletti-C. Brass-L-F. TI Proteolysis of the human platelet and endothelial cell thrombin receptor by neutrophil-derived cathepsin G SO J-Biol-Chem. 1995 May 12. 270(19). p 11168. AU Janssen-J-J-M. Bovee-Geurts-P-H-M. Merkx-M. DeGrip-W-J. TI Histidine tagging both allows convenient single-step purification of bovine rhodopsin and exerts ionic strength-dependent effects on its photochemistry SO J-Biol-Chem. 1995 May 12. 270(19). p 11222. AU Inoue-I. Rohrwasser-A. Helin-C. Jeunemaitre-X. Crain-P. Bohlender-J. Lifton-R-P. Corvol-P. Ward-K. Lalouel-J-M. TI A mutation of angiotensinogen in a patient with preeclampsia leads to altered kinetics of the renin-angiotensin system SO J-Biol-Chem. 1995 May 12. 270(19). p 11430. AU Hesselgesser-J. Chitnis-C-E. Miller-L-H. Yansura-D-G. Simmons-L-C. Fairbrother-W-J. Kotts-C. Wirth-C. Gillece-Castro-B-L. Horuk-R. TI A mutant of melanoma growth stimulating activity does not activate neutrophils but blocks erythrocyte invasion by malaria SO J-Biol-Chem. 1995 May 12. 270(19). p 11472. AU Wu-J. Spiegel-S. Sturgill-T-W. TI Sphingosine 1-phosphate rapidly activates the mitogen-activated protein kinase pathway by a G protein-dependent mechanism SO J-Biol-Chem. 1995 May 12. 270(19). p 11484. AU Hellevuo-K. Yoshimura-M. Mons-N. Hoffman-P-L. Cooper-D-M-F. Tabakoff-B. TI The characterization of a novel human adenylyl cyclase which is present in brain and other tissues SO J-Biol-Chem. 1995 May 12. 270(19). p 11581. From owner-7tms_r@net.bio.net Wed May 03 23:00:00 1995 Path: biosci!COMPUSERVE.COM!74757.3155 From: 74757.3155@COMPUSERVE.COM (Ryosuke Furubayashi) Newsgroups: bionet.molbio.proteins.7tms_r Subject: HELP Date: 4 May 1995 12:45:33 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 2 Sender: daemon@net.bio.net Distribution: world Message-ID: <950504194341_74757.3155_EHL194-2@CompuServe.COM> NNTP-Posting-Host: net.bio.net RYOSUKE FURUBAYASHI From owner-7tms_r@net.bio.net Thu May 04 23:00:00 1995 Path: biosci!PT.CYANAMID.COM!hadcockj From: hadcockj@PT.CYANAMID.COM Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re[2]: HEK 293 cells and receptors Date: 5 May 1995 06:41:45 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HQ5145DM0IBK1APN@ptag2.pt.Cyanamid.COM> NNTP-Posting-Host: net.bio.net Terry Reisine reported that HEK cells express low levels (20 fmol/mg) of somatostatin receptor subtype 2. John Hadcock hadcockj@pt.cyanamid.com ______________________________ Reply Separator _________________________________ Subject: Re: HEK 293 cells and receptors Author: boundy@SYNAPSE.BMS.COM at GATEWAY Date: 5/4/95 1:02 PM endogenous 7TM receptors in HEK 293 cells.. Sorry, I missed the word endogenous before... D1 dopamine receptors are endogenously expressed at realtively low densities in HEK 293 cells. They do couple to stimulation of adenylyl cyclase in our hands -*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*-*- Virginia Boundy Dept of Pharmacology University of Pennsylvania Philadelphia, PA 19104 -- CNS Exploratory Bristol-Myers Squibb Pharmaceutical Research Institute Wallingford, CT 06492 (203) 284-6927 From owner-7tms_r@net.bio.net Fri May 05 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!zib-berlin.de!cs.tu-berlin.de!fauern!winx03!wrzx15!phak004 From: phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: rhodopsin coordinates Date: 5 May 1995 08:51:56 GMT Organization: University of Wuerzburg, Germany Lines: 14 Message-ID: <3ocovc$dur@winx03.informatik.uni-wuerzburg.de> References: <3o8jjl$1ko@usenet.INS.CWRU.Edu> NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] Bryan Roth (roth@biocserver.cwru.edu) wrote: > Some time ago, Baldwin published a model of rhodopsin based on her > 2-D crystals. Does anyone know how of if the coordinates of this > model may be obtained. AFAIK the publication of the model was only announced in an abstract (can't remember which conference). I haven't seen an original publication yet. --Cornelius. -- /* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ /* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Fri May 05 23:00:00 1995 Path: biosci!agate!howland.reston.ans.net!Germany.EU.net!zib-berlin.de!cs.tu-berlin.de!fauern!winx03!wrzx15!phak004 From: phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Crystals? Date: 5 May 1995 08:54:51 GMT Organization: University of Wuerzburg, Germany Lines: 17 Message-ID: <3ocp4r$dur@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] I have heard a rumour recently that Brian Kobilka has been able to grow crystals from beta_2-adrenergic receptors. Can anyone confirm/deny this? On a related topic, I remember Rick Neubig asking for news about G protein crystallization some time ago, and I think he didn't get an answer. Well, I had the opportunity to attend a talk of Alfred Gilman recently, and he confirmed that they have grown crystals from alpha-beta-gamma heterotrimeric G proteins as well as crystals from beta-gamma subunits. They have tried to solve the structure of the heterotrimeric crystals by molecular replace- ment, but unfortunately they didn't succeed. --Cornelius. -- /* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ /* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ /* "Science is the game you play with God to find out what His rules are." */ From owner-7tms_r@net.bio.net Sun May 07 23:00:00 1995 Path: biosci!WELCHLINK.WELCH.JHU.EDU!kzierler From: kzierler@WELCHLINK.WELCH.JHU.EDU (KENNETH ZIERLER) Newsgroups: bionet.molbio.proteins.7tms_r Subject: G-protein Date: 8 May 1995 08:42:15 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 4 Sender: daemon@net.bio.net Distribution: world Message-ID: NNTP-Posting-Host: net.bio.net Michael Levine tells me you have a receptor-G-protein bulletin board. I'd like to subscribe, please. Kenneth Zierler, medicine & physiology,Johns Hopkins Med kzierler@welchlink.welch.jhu.edu From owner-7tms_r@net.bio.net Sun May 07 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!tgevax.life.uiuc.edu!saurav From: saurav@tgevax.life.uiuc.edu (Saurav Misra) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: rhodopsin coordinates Date: 8 May 95 17:56:00 GMT Organization: University of Illinois at Urbana Lines: 26 Message-ID: References: <3o8jjl$1ko@usenet.INS.CWRU.Edu> <3ocovc$dur@winx03.informatik.uni-wuerzburg.de> NNTP-Posting-Host: tgevax.life.uiuc.edu phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) writes: >Bryan Roth (roth@biocserver.cwru.edu) wrote: >> Some time ago, Baldwin published a model of rhodopsin based on her >> 2-D crystals. Does anyone know how of if the coordinates of this >> model may be obtained. >AFAIK the publication of the model was only announced in an abstract (can't >remember which conference). I haven't seen an original publication yet. >--Cornelius. >-- >/* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ >/* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ >/* "Science is the game you play with God to find out what His rules are." */ The latest issue of Biophysical Journal (May,v.68) has an article on the latest diffraction structure obtained by Schertler's group. I know they work with Richard Henderson and Baldwin (and, as a matter of fact, use Baldwin's aid in their structure analysis; her model, that is). -Saurav Misra saurav@tgevax.life.uiuc.edu U. of Illinois Biophysics Dept. From owner-7tms_r@net.bio.net Sun May 07 23:00:00 1995 Path: biosci!rutgers!uwm.edu!psuvax1!news.ecn.bgu.edu!usenet.ins.cwru.edu!usenet From: Bryan Roth Newsgroups: bionet.molbio.proteins.7tms_r Subject: re: rhodopsin Date: 8 May 1995 21:26:14 GMT Organization: Case Western Reserve University, Cleveland, Ohio (USA) Lines: 7 Message-ID: <3om29m$qua@usenet.INS.CWRU.Edu> NNTP-Posting-Host: meds20785.meds.cwru.edu From Suarav Misra: >The latest issue of Biophysical Journal (May,v.68) has an article on the latest diffraction structure obtained by Schertler's group. Other than that there is apparantly nothing new that is public. Anything else of interest I will pass along From owner-7tms_r@net.bio.net Sun May 07 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bloom-beacon.mit.edu!uhog.mit.edu!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!uunet!in1.uu.net!hearst.acc.Virginia.EDU!murdoch!usenet From: "Bruce D. Gaylinn" Subject: Re: HEK 293 cells and receptors X-Nntp-Posting-Host: bootp-205-42.bootp.virginia.edu Content-Type: text/plain; charset=us-ascii Message-ID: Sender: usenet@murdoch.acc.Virginia.EDU Content-Transfer-Encoding: 7bit Organization: University of Virginia References: <9505040635.AA11220@nngate.novo.dk> Mime-Version: 1.0 Date: Mon, 8 May 1995 15:58:08 GMT X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Lines: 31 bsw@novo.dk (Birgitte Schjellerup Wulff) wrote: > Hi > >Have anyone experience with endogenous 7TM receptors in HEK 293 cells. I'm >especially interested in knowing if receptors from the secretin/VIP >receptor family is endogenous expressed in the cells. > >Best regards > >Birgitte Wulff > >Birgitte Wulff >bsw@novo.dk< >Dept. of Molecular Pharmacology 1C.S36, Pharmaceutical Biotechnology >Novo-Nordisk >2880 Bagsvaerd, Denmark >tlf: +45 44 42 34 21 fax: +45 44 98 50 07 > > Hi. We have been working with GHRH receptor expressed in HEK293s. The non-transfected control 293s do show traces of GHRH binding that is inhibited by GTP. A corresponding cAMP response to GHRH is also seen. These responses are weak and suggestive of a low affinity site. No trace of GHRH receptor message is detected on a northern. Though we have not examined it in detail, we assume that the GHRH is acting through an endogenous secretin family receptor on the HEK293s. Could be VIP. Bruce Gaylinn Univ. of Virginia From owner-7tms_r@net.bio.net Mon May 08 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!bloom-beacon.mit.edu!spool.mu.edu!howland.reston.ans.net!pipex!warwick!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (RM. Eustace) Subject: 32p NAD labelling by pertussis toxin Message-ID: Sender: usenet@info.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: zeus.bris.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL2] Date: Tue, 9 May 1995 12:26:17 GMT Lines: 21 Dear All I am trying to establish a link between receptor and G-protein activation. One method that I've tried is pertussis toxin mediated ribosylation of susceptible G-proteins in the presence of 32p NAD, thus specifically tagging the G-protein prior to SDS-PAGE. This works well. I have also tried to pretreat the whole cells with pertussis toxin first in order to abolish the signal following a second round of "hot" ribosylation. This also works well and in a time dependent manner (the siganl is abolished by 6 hrs PTX pretreatment. The problem is that as the signal disappears, a second, lighter band appears. I cannot rid myself of this band even with extensive PTX pretreatment regimes. The reason that I want to get rid of this band is that I have come to realize that PTX -mediated ribosylation is extremely specific, therefore this second band must be a G-alpha subunit of some description and cannot therefore be ignored. The eventual aim is to preincubate with pertussis AND the ligand of interest to restore the signal (the first round of ribosylation would be only partially successful since petussis only labels the "resting" alpha subunit. Thankyou in advance Adam Collier p.s. I am aware that there are many other methods of analysing G-proteins!!!! From owner-7tms_r@net.bio.net Tue May 09 23:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!noc.netcom.net!usenet.eel.ufl.edu!spool.mu.edu!uwm.edu!psuvax1!news.pop.psu.edu!news.cac.psu.edu!newsserver.jvnc.net!netnews.sb.com!news From: Steven_Mcclue-1%notes@sb.com (Steve Mcclue) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: 32p NAD labelling by pertussis toxin Date: 10 May 1995 08:31:00 GMT Organization: Smithkline Beecham Pharmaceuticals Lines: 23 Message-ID: <3optk4$pko@phunn1.sb.com> References: NNTP-Posting-Host: had542.uk.sb.com Mime-Version: 1.0 X-Newsreader: WinVN 0.99.2 In article , dhajc@zeus.bris.ac.uk says... >I have also tried to pretreat the whole cells with pertussis toxin first in >order to abolish the signal following a second round of "hot" >ribosylation. This also works well and in a time dependent manner (the >siganl is abolished by 6 hrs PTX pretreatment. The problem is that as >the signal disappears, a second, lighter band appears. I cannot rid >myself of this band even with extensive PTX pretreatment regimes. >Thankyou in advance >Adam Collier >p.s. I am aware that there are many other methods of analysing G-proteins!!!! You don't say what the MW of this phantom band is - that might help us to identify it. If you do your experiment with a control consisting of everything *except* pertussis toxin (ie ribosylate a sample of your membranes using only toxin vehicle - DTT or whatever you use), do you still have a band at this position, or does it only appear in the presence of PT ? There is a nice chapter in Methods in Enzymol vol 195 pp257-266 (Kopf and Woolkalis) which discusses some of the problems associated with pertussis toxin ribosylation. Of course, at the end of the day, you might have found a new G-protein! Steve McClue From owner-7tms_r@net.bio.net Tue May 09 23:00:00 1995 Path: biosci!agate!news.ucdavis.edu!library.ucla.edu!psgrain!iafrica.com!ticsa.com!cstatd.cstat.co.za!ucthpx!chempath.uct.ac.za!james From: james@chempath.uct.ac.za (James Davidson) Newsgroups: bionet.molbio.proteins.7tms_r Subject: First intracellular loop basic residues Date: Wed, 10 May 1995 14:04:06 GMT Organization: University of Cape Town Lines: 6 Message-ID: NNTP-Posting-Host: 137.158.135.10 Does anyone have any information about the role of basic residues in the first intracellular loop of GPCRs? I would be most grateful for any help on this topic. James Davidson, Cape Town From owner-7tms_r@net.bio.net Tue May 09 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!daresbury!sunsite.doc.ic.ac.uk!lyra.csx.cam.ac.uk!warwick!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (AJ. Collier) Subject: high molecular weight G-proteins Message-ID: Sender: usenet@info.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: zeus.bris.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL2] Date: Wed, 10 May 1995 10:44:33 GMT Lines: 7 Dear All, I have read a single paper regarding a heterotrimeric G-protein with a mass of around 67kDa. Has this been verified?, are there any details in other papers? Thanks Adam Collier From owner-7tms_r@net.bio.net Wed May 10 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!xlink.net!rz.uni-karlsruhe.de!news.uni-stuttgart.de!uni-regensburg.de!fauern!winx03!wrzx15!phak004 From: phak004@wrzx15.rz.uni-wuerzburg.de (Cornelius Krasel) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: high molecular weight G-proteins Date: 11 May 1995 12:53:58 GMT Organization: University of Wuerzburg, Germany Lines: 71 Message-ID: <3ot1d6$4nu@winx03.informatik.uni-wuerzburg.de> References: NNTP-Posting-Host: wrzx15.rz.uni-wuerzburg.de X-Newsreader: TIN [version 1.2 PL2] AJ. Collier (dhajc@zeus.bris.ac.uk) wrote: > I have read a single paper regarding a heterotrimeric G-protein with a > mass of around 67kDa. Has this been verified?, are there any details in > other papers? To which paper do you refer? Two come to mind: @article{kehlenbach:94, author = {Ralph H. Kehlenbach and Julia Matthey and Wieland B. Huttner}, title = {{XL}$\alpha$s is a new type of {G} protein.}, journal = {Nature}, volume = 372, pages = {804--809}, year = 1994 } (I haven't seen any other publications on this) or @article{nakaoka:94, author = {Hideaki Nakaoka and Dianne M. Perez and Kwang Jin Baek and Tanya Das and Ahsan Husain and Kunio Misono and Mie-Jae Im and Robert M. Graham}, title = {{G}$_{\rm h}$: A {GTP}-Binding Protein with Transglutaminase Activity and Receptor Signaling Function.}, journal = {Science}, volume = 264, pages = {1593--1596}, year = 1994 } The existence of the latter protein has been inferred previously: @article{baek:93, author = {Kwang Jin Baek and Tanya Das and Caroline Gray and Shareef Antar and Gurunathan Murugesan and Mie-Jae Im}, title = {Evidence That the {G}$_{\rm h}$ Protein Is a Signal Mediator from $\alpha_1$-Adrenoceptor to a Phospholipase {C}. {I. I}dentification of $\alpha_1$-adrenoceptor-coupled {G}$_{\rm h}$ family and purification of {G}$_{\rm h7}$ from bovine heart.}, journal = JBC, volume = 268, number = 36, pages = {27390--27397}, year = 1993 } @article{das:93, author = {Tanya Das and Kwang Jin Baek and Caroline Gray and Mie-Jae Im}, title = {Evidence That the {G}$_{\rm h}$ Protein Is a Signal Mediator from $\alpha_1$-Adrenoceptor to a Phospholipase {C}. {II. P}urification and characterization of a {G}$_{\rm h}$-coupled 69-{kDa} phospholipase {C} and reconstitution of $\alpha_1$-adrenoceptor, {G}$_{\rm h}$ family, and phospholipase {C}.}, journal = JBC, volume = 268, number = 36, pages = {27398--27405}, year = 1993 } --Cornelius. -- /* Cornelius Krasel, Institut f. Pharmakologie, Versbacher Str. 9, D-97078 */ /* Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de */ /* "Science is the game you play with God to find out what His rules are." */ /* --Annoy a Scientologist: contribute to the Dennis Erlich Defence Fund-- */ From owner-7tms_r@net.bio.net Thu May 11 23:00:00 1995 Path: biosci!adam.cc.sunysb.edu!news.sprintlink.net!howland.reston.ans.net!usenet.ins.cwru.edu!usenet From: Bryan Roth Newsgroups: bionet.molbio.proteins.7tms_r Subject: Receptor purification position Date: 12 May 1995 18:48:55 GMT Organization: Case Western Reserve University, Cleveland, Ohio (USA) Lines: 21 Message-ID: <3p0ain$e5g@usenet.INS.CWRU.Edu> NNTP-Posting-Host: meds20785.meds.cwru.edu A post-doctoral position is availible in my laboratory starting 1 July 1995 or later. We currently have the available technology to purify large quantities of a G-protein coupled receptor via histidine-"tags". Via collaboration with biophysicists and crystallographers in the Department of Biochemistry we are devoting considerable effort to begin to elucidate the structure of a model serotonin receptor. The position will be for at least two years with possible extension beyond this point. Expertise in protein purification essential. If you are interested in a project which could have a major impact on the field, please contact me at: Bryan Roth roth@biocserver.cwru.edu Department of Biochemistry Case Western Reserve University School of Medicine Room W 438 11100 Euclid Avenue Cleveland, OH 44106-4935 From owner-7tms_r@net.bio.net Sat May 13 23:00:00 1995 Path: biosci!RECEPTOR.MGH.HARVARD.EDU!lfk From: lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: TOCs of interest to 7tms_r JBC_May_19 Date: 14 May 1995 16:53:22 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505142346.AA04858@receptor> NNTP-Posting-Host: net.bio.net AU Ben-Baruch-A. Michiel-D-F. Oppenheim-J-J. TI Signals and receptors involved in recruitment of inflammatory cells SO J-Biol-Chem. 1995 May 19. 270(20). p 11703. AU Pitcher-J-A. Touhara-K. Payne-E-S. Lefkowitz-R-J. TI Pleckstrin homology domain-mediated membrane association and activation of the {beta}-adrenergic receptor kinase requires coordinate interaction with G{beta}{gamma} subunits and lipid SO J-Biol-Chem. 1995 May 19. 270(20). p 11707. AU Balboa-M-A. Balsinde-J. Dennis-E-A. Insel-P-A. TI A phospholipase D-mediated pathway for generating diacylglycerol in nuclei from Madin-Darby canine kidney cells SO J-Biol-Chem. 1995 May 19. 270(20). p 11738. AU Selbie-L-A. Darby-K. Schmitz-Peiffer-C. Browne-C-L. Herzog-H. Shine-J. Biden-T-J. TI Synergistic interaction of Y1-neuropeptide Y and {alpha}1b-adrenergic receptors in the regulation of phospholipase C, protein kinase C, and arachidonic acid production SO J-Biol-Chem. 1995 May 19. 270(20). p 11789. AU James-S-R. Paterson-A. Harden-T-K. Downes-C-P. TI Kinetic analysis of phospholipase C{beta} isoforms using phospholipid-detergent mixed micelles. Evidence for interfacial catalysis involving distinct micelle binding and catalytic steps. SO J-Biol-Chem. 1995 May 19. 270(20). p 11872. From owner-7tms_r@net.bio.net Tue May 16 23:00:00 1995 Path: biosci!ORION.RZ.MDC-BERLIN.DE!mhoehe From: mhoehe@ORION.RZ.MDC-BERLIN.DE (Margret Hoehe) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Graduate Student Position Available for DNA Sequence Analysis in G Protein-Coupled Receptor Genes Date: 17 May 1995 01:17:11 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 29 Sender: daemon@net.bio.net Distribution: world Message-ID: <199505170815.KAA11363@orion.rz.mdc-berlin.de> NNTP-Posting-Host: net.bio.net The 'Genome Research' working group at the Max Delbrueck Center for Molecular Medicine in Berlin-Buch is looking for a Graduate Student (Doktorand/in) (BAT-O IIa/2). We have been developing (in close collaboration with Dr. George Church, Harvard Medical School) a 'Multiplex PCR Sequencing' approach to the large scale analysis of interindividual DNA sequence variation in G protein-coupled receptor genes and other biomedically important target regions. We will continue this line of technology development, which allows us at present to sequence 20 PCR fragments simultaneously, and we plan to evaluate sets of G protein-coupled receptor genes in healthy and diseased individuals. The position is available now and will be for at least two years. Expertise in PCR, sequencing and possibly biocomputing wanted. Please contact Margret Hoehe. Margret Hoehe Max-Delbrueck-Center for Molecular Medicine Robert-Roessle-Str. 10 D-13122 Berlin Phone (030) 9406 3154 (office) (030) 9406 3312 (lab) Fax (030) 9406 3842 From owner-7tms_r@net.bio.net Thu May 18 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: wzhao80850@aol.com (WZhao80850) Newsgroups: bionet.molbio.proteins.7tms_r Subject: HELP needed in western! Date: 19 May 1995 16:31:07 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 15 Sender: root@newsbf02.news.aol.com Message-ID: <3piv6b$p2h@newsbf02.news.aol.com> Reply-To: wzhao80850@aol.com (WZhao80850) NNTP-Posting-Host: newsbf02.mail.aol.com Hi there, I have experienced high background problem in my western blotting. I am using SDS-PAGE in non-reduced condition. The primary antibody dilution is pretty high, i.e. 1:25 (10 ug/ml) in TBST and 1% milk. Is there any way I may reduce the background? Also, the protein bands are bended and curved on the film. Why is that? Any suggestion will helpful for me, thanks in advance! Cheers! Wei Wei Zhao WZhao80850@aol.com or weiz@protein.bchs.uh.edu Tel: (713)669-1091(H), (713)790-4696 Baylor College of Medicine Houston, TX 77030 From owner-7tms_r@net.bio.net Fri May 19 23:00:00 1995 Path: biosci!BIOMED.MED.YALE.EDU!KARNE From: KARNE@BIOMED.MED.YALE.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: (none) Date: 20 May 1995 18:09:20 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 2 Sender: daemon@net.bio.net Distribution: world Message-ID: <01HQQ6IEGA4M00B2UE@BIOMED.MED.YALE.EDU> NNTP-Posting-Host: net.bio.net unsubscribe From owner-7tms_r@net.bio.net Sun May 21 23:00:00 1995 Path: biosci!daresbury!not-for-mail From: Newsgroups: bionet.molbio.proteins.7tms_r Subject: plc-betas Date: 22 May 1995 18:05:36 +0100 Lines: 10 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3pqg90$dse@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Hi, does anyone know of recent articles (or even better a review) on the tissue specificity of expression of phospholipase C-beta isozymes? thanks, Arturo Galvani Pharmacia Milan From owner-7tms_r@net.bio.net Sun May 21 23:00:00 1995 Newsgroups: bionet.molbio.proteins.7tms_r Path: biosci!agate!sunsite.doc.ic.ac.uk!nntp0.brunel.ac.uk!strath-cs!dcl-cs!gdt!bsmail!zeus!dhajc From: dhajc@zeus.bris.ac.uk (HE. Fergus) Subject: Re: HELP needed in western! Message-ID: Sender: usenet@info.bris.ac.uk (Usenet news owner) Nntp-Posting-Host: zeus.bris.ac.uk Organization: University of Bristol, England X-Newsreader: TIN [version 1.2 PL2] References: <3piv6b$p2h@newsbf02.news.aol.com> Date: Mon, 22 May 1995 09:02:43 GMT Lines: 22 WZhao80850 (wzhao80850@aol.com) wrote: : Hi there, : I have experienced high background problem in my western blotting. I am : using SDS-PAGE in non-reduced condition. The primary antibody dilution is : pretty high, i.e. 1:25 (10 ug/ml) in TBST and 1% milk. Is there any way I : may reduce the background? Also, the protein bands are bended and curved : on the film. Why is that? : Any suggestion will helpful for me, thanks in advance! : Cheers! : Wei : Wei Zhao : WZhao80850@aol.com or weiz@protein.bchs.uh.edu : Tel: (713)669-1091(H), (713)790-4696 : Baylor College of Medicine : Houston, TX 77030 I found that replacing tween with Triton x100 improved my G-protein western blots a lot. Also does the antibody work at a lower concentration, say 1ug/ml. The protein bands may well be curved coz you are running the gel too hot. From owner-7tms_r@net.bio.net Mon May 22 23:00:00 1995 Path: biosci!rutgers!netnews.upenn.edu!cronkite.ocis.temple.edu!VM.TEMPLE.EDU!SHICKLEY From: SHICKLEY@VM.TEMPLE.EDU Newsgroups: bionet.molbio.proteins.7tms_r Subject: PIR gopher or ftp site? Date: Tue, 23 May 95 09:44:44 EDT Organization: Temple University Lines: 10 Message-ID: <173A7890CS86.SHICKLEY@VM.TEMPLE.EDU> NNTP-Posting-Host: vm.temple.edu I lost my PIR Newletter so I don't have this info. Does anyone know of an ftp site or gopher site for the Protein Identification Resource? Any help would be appreciated. You can post here. TIM SHICKLEY SHICKLEY@VM.TEMPLE.EDU TSHICKLEY@HAL.DENTAL.TEMPLE.EDU shickley@astro.ocis.temple.edu -----------------------STANDARD DISCLAIMERS APPLY----------------- From owner-7tms_r@net.bio.net Tue May 23 23:00:00 1995 Path: biosci!MACE.CC.PURDUE.EDU!kmartin From: kmartin@MACE.CC.PURDUE.EDU (katherine) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: photolabeling azidoGTP Date: 24 May 1995 15:19:03 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 15 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505242218.AA02968@mace.cc.purdue.edu> NNTP-Posting-Host: net.bio.net I personally haven't seen any papers dealing with the use of Azidoanilido- GTP in a permeabilized cell system. Unfortunately I lost one of the messages that discussed the use of azidoanilido-GTP for labelling thus some of the references I give you might be repeats. Analytical Biochemistry 214, 171-178(1993) Biochemistry 24, 1555-1560(1985) Methods Enzymology vol. 195 I hope this helps. Katherine From owner-7tms_r@net.bio.net Tue May 23 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!swrinde!emory!news-feed-1.peachnet.edu!news.duke.edu!altschul From: altschul@PROBLEM_WITH_INEWS_GATEWAY_FILE (Danny Altshuler) Newsgroups: bionet.molbio.proteins.7tms_r Subject: photolabeling azidoGTP Date: 24 May 1995 19:26:18 GMT Organization: Duke University, Durham, NC, USA Lines: 8 Message-ID: <3q018q$r3r@news.duke.edu> NNTP-Posting-Host: abacus.mc.duke.edu X-Newsreader: TIN [version 1.2 PL2] Does anyone have expierence with 8-azidoGTP (alphaP32) on permeabilized cells? Can you do labeling by permeabilization and then agonist stimulation to follow the specific agonist-stimulated G-protein? How good or bad is the background? Any comercially available azido? Please if anyone have any protocols, references or any kind of help pleas e-mail me to altschul@abacus.mc.duke.edu Thanks, Danny From owner-7tms_r@net.bio.net Tue May 23 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!newsrelay.iastate.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Selectivity in alpha1b AR i3 regions Date: 24 May 1995 16:56:04 GMT Organization: University of Michigan Lines: 21 Message-ID: <3pvof5$53k@lastactionhero.rs.itd.umich.edu> NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1b3 (Windows; I; 16bit) There is an interesting paper by Wu, Jiang and Simon in JBC (270:9828-9832, 1995) showing distinct regions of the alpha1b AR i3 loop were required for coupling to alpha-q/11 vs alpha-14 vs alpha-16. Was anyone else bothered by the fact that most mutations knocked out q/11 responses, a few more knocked out alpha-14 responses and only one knocked out alpha-16? The sensitivity to mutation was inversely correlated with the amount of G protein alpha subunit! There is a very small amount of q/11 (endogenous), more alpha-14 and more alpha-16? Is this really specificity or just progressive crippling of a receptor so that it needs more and more G protein to mediate the response? Seems like a more quantitative analysis would be required. _________________________________________________________ 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 May 24 23:00:00 1995 Path: biosci!RECEPTOR.MGH.HARVARD.EDU!lfk From: lfk@RECEPTOR.MGH.HARVARD.EDU (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: JBC TOC for 7tms_r 5/26 and 6/2 Date: 25 May 1995 10:20:57 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 81 Sender: daemon@net.bio.net Distribution: world Message-ID: <9505251216.AA01403@receptor> NNTP-Posting-Host: net.bio.net AU Liu-W. Yoon-J. Burg-M. Chen-L. Pak-W-L. TI Molecular characterization of two Drosophila guanylate cyclases expressed in the nervous system SO J-Biol-Chem. 1995 May 26. 270(21). p 12418. AU Linseman-D-A. Benjamin-C-W. Jones-D-A. TI Convergence of angiotensin II and platelet-derived growth factor receptor signaling cascades in vascular smooth muscle cells SO J-Biol-Chem. 1995 May 26. 270(21). p 12563. AU Deer-J-L-R. Hurley-J-B. Yarfitz-S-L. TI G protein control of Drosophila photoreceptor phospholipase C SO J-Biol-Chem. 1995 May 26. 270(21). p 12623. AU Meng-F. Hoversten-M-T. Thompson-R-C. Taylor-L. Watson-S-J. Akil-H. TI A chimeric study of the molecular basis of affinity and selectivity of the {kappa} and the {delta} opioid receptors. Potential role of extracellular domains. SO J-Biol-Chem. 1995 May 26. 270(21). p 12730. AU Tholanikunnel-B-G. Granneman-J-G. Malbon-C-C. TI The Mr 35,000 {beta}-adrenergic receptor mRNA-binding protein binds transcripts of G-protein-linked receptors which undergo agonist-induced destabilization SO J-Biol-Chem. 1995 May 26. 270(21). p 12787. AU Feng-Y-H. Noda-K. Saad-Y. Liu-X-. Husain-A. Karnik-S-S. TI The docking of Arg2 of angiotensin II with Asp281 of AT1 receptor is essential for full agonism SO J-Biol-Chem. 1995 May 26. 270(21). p 12846. AU Konrad-R-J. Young-R-A. Record-R-D. Smith-R-M. Butkerait-P. Manning-D. Jarett-L. Wolf-B-A. TI The heterotrimeric G-protein Gi is localized to the insulin secretory granules of {beta}-cells and is involved in insulin exocytosis SO J-Biol-Chem. 1995 May 26. 270(21). p 12869. AU Garrett-J-E. Capuano-I-V. Hammerland-L-G. Hung-B-C-P. Brown-E-M. Hebert-S-C. Nemeth-E-F. Fuller-F. TI Molecular cloning and functional expression of human parathyroid calcium receptor cDNAs SO J-Biol-Chem. 1995 May 26. 270(21). p 12919. AU Moolenaar-W-H. TI Lysophosphatidic acid, a multifunctional phospholipid messenger SO J-Biol-Chem. 1995 June 2. 270(22). p 12949. AU Xue-J-C. Chen-C. Zhu-J. Kunapuli-S-P. Riel-J-K-. Yu-L. Liu-Chen-L-Y. TI The third extracellular loop of the {mu} opioid receptor is important for agonist selectivity SO J-Biol-Chem. 1995 June 2. 270(22). p 12977. AU Luttrell-L-M. Hawes-B-E. Touhara-K. Biesen-T-. Koch-W-J. Lefkowitz-R-J. TI Effect of cellular expression of pleckstrin homology domains on Gi-coupled receptor signaling SO J-Biol-Chem. 1995 June 2. 270(22). p 12984. AU Skiba-N-P. Artemyev-N-O. Hamm-H-E. TI The carboxyl terminus of the {gamma}-subunit of rod cGMP phosphodiesterase contains distinct sites of interaction with the enzyme catalytic subunits and the {alpha}-subunit of transducin SO J-Biol-Chem. 1995 June 2. 270(22). p 13210. AU McKay-R-R. Chen-D-M. Miller-K. Kim-S. Stark-W-S. Shortridge-R-D. TI Phospholipase C rescues visual defect in norpA mutant of Drosophila melanogaster SO J-Biol-Chem. 1995 June 2. 270(22). p 13271. AU Soltoff-S-P. Toker-A. TI Carbachol, substance P, and phorbol ester promote the tyrosine phosphorylation of protein kinase C{delta} in salivary gland epithelial cells SO J-Biol-Chem. 1995 June 2. 270(22). p 13490. From owner-7tms_r@net.bio.net Wed May 24 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!lamarck.sura.net!news.uky.edu!chuck From: Chuck Staben Newsgroups: bionet.molbio.proteins.7tms_r Subject: fungal g-proteins Date: 24 May 1995 23:30:15 GMT Organization: University of Kentucky Computing Services Lines: 7 Message-ID: <3q0fi7$rmn@service1.uky.edu> NNTP-Posting-Host: bs6.mi.uky.edu I am interested in finding chemical inhibitors known to act on fungal G-proteins, either agonists or antagonists. I am currently working with a fungal G-protein system that cannot readily be manipulated by genetics, so I would like to observe the effects that a G-protein system is having by applying chemicals to intact cells and monitoring physiological responses. From owner-7tms_r@net.bio.net Wed May 24 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!vixen.cso.uiuc.edu!uwm.edu!rutgers!oitnews.harvard.edu!bloch.nmr.mgh.harvard.edu!nntp.mgh.harvard.edu!lfk From: lfk@receptor.mgh.harvard.edu (Lee F. (Frank) Kolakowski) Newsgroups: bionet.molbio.proteins.7tms_r Subject: JBC TOC for 7tms_r 5/26 and 6/2 Date: 25 May 1995 12:13:02 GMT Organization: Mass. General Hospital, Renal Unit Lines: 85 Distribution: world Message-ID: NNTP-Posting-Host: receptor.mgh.harvard.edu AU Liu-W. Yoon-J. Burg-M. Chen-L. Pak-W-L. TI Molecular characterization of two Drosophila guanylate cyclases expressed in the nervous system SO J-Biol-Chem. 1995 May 26. 270(21). p 12418. AU Linseman-D-A. Benjamin-C-W. Jones-D-A. TI Convergence of angiotensin II and platelet-derived growth factor receptor signaling cascades in vascular smooth muscle cells SO J-Biol-Chem. 1995 May 26. 270(21). p 12563. AU Deer-J-L-R. Hurley-J-B. Yarfitz-S-L. TI G protein control of Drosophila photoreceptor phospholipase C SO J-Biol-Chem. 1995 May 26. 270(21). p 12623. AU Meng-F. Hoversten-M-T. Thompson-R-C. Taylor-L. Watson-S-J. Akil-H. TI A chimeric study of the molecular basis of affinity and selectivity of the {kappa} and the {delta} opioid receptors. Potential role of extracellular domains. SO J-Biol-Chem. 1995 May 26. 270(21). p 12730. AU Tholanikunnel-B-G. Granneman-J-G. Malbon-C-C. TI The Mr 35,000 {beta}-adrenergic receptor mRNA-binding protein binds transcripts of G-protein-linked receptors which undergo agonist-induced destabilization SO J-Biol-Chem. 1995 May 26. 270(21). p 12787. AU Feng-Y-H. Noda-K. Saad-Y. Liu-X-. Husain-A. Karnik-S-S. TI The docking of Arg2 of angiotensin II with Asp281 of AT1 receptor is essential for full agonism SO J-Biol-Chem. 1995 May 26. 270(21). p 12846. AU Konrad-R-J. Young-R-A. Record-R-D. Smith-R-M. Butkerait-P. Manning-D. Jarett-L. Wolf-B-A. TI The heterotrimeric G-protein Gi is localized to the insulin secretory granules of {beta}-cells and is involved in insulin exocytosis SO J-Biol-Chem. 1995 May 26. 270(21). p 12869. AU Garrett-J-E. Capuano-I-V. Hammerland-L-G. Hung-B-C-P. Brown-E-M. Hebert-S-C. Nemeth-E-F. Fuller-F. TI Molecular cloning and functional expression of human parathyroid calcium receptor cDNAs SO J-Biol-Chem. 1995 May 26. 270(21). p 12919. AU Moolenaar-W-H. TI Lysophosphatidic acid, a multifunctional phospholipid messenger SO J-Biol-Chem. 1995 June 2. 270(22). p 12949. AU Xue-J-C. Chen-C. Zhu-J. Kunapuli-S-P. Riel-J-K-. Yu-L. Liu-Chen-L-Y. TI The third extracellular loop of the {mu} opioid receptor is important for agonist selectivity SO J-Biol-Chem. 1995 June 2. 270(22). p 12977. AU Luttrell-L-M. Hawes-B-E. Touhara-K. Biesen-T-. Koch-W-J. Lefkowitz-R-J. TI Effect of cellular expression of pleckstrin homology domains on Gi-coupled receptor signaling SO J-Biol-Chem. 1995 June 2. 270(22). p 12984. AU Skiba-N-P. Artemyev-N-O. Hamm-H-E. TI The carboxyl terminus of the {gamma}-subunit of rod cGMP phosphodiesterase contains distinct sites of interaction with the enzyme catalytic subunits and the {alpha}-subunit of transducin SO J-Biol-Chem. 1995 June 2. 270(22). p 13210. AU McKay-R-R. Chen-D-M. Miller-K. Kim-S. Stark-W-S. Shortridge-R-D. TI Phospholipase C rescues visual defect in norpA mutant of Drosophila melanogaster SO J-Biol-Chem. 1995 June 2. 270(22). p 13271. AU Soltoff-S-P. Toker-A. TI Carbachol, substance P, and phorbol ester promote the tyrosine phosphorylation of protein kinase C{delta} in salivary gland epithelial cells SO J-Biol-Chem. 1995 June 2. 270(22). p 13490. -- Frank Kolakowski Email: lfk@receptor.mgh.harvard.edu 617-355-7515 (LAB) GCRDb-WWW From owner-7tms_r@net.bio.net Wed May 24 23:00:00 1995 Path: biosci!rutgers!uwm.edu!vixen.cso.uiuc.edu!howland.reston.ans.net!news-e1a.megaweb.com!newstf01.news.aol.com!newsbf02.news.aol.com!not-for-mail From: microplate@aol.com (MICROPLATE) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Microtiter 96 well Date: 25 May 1995 20:09:22 -0400 Organization: America Online, Inc. (1-800-827-6364) Lines: 7 Sender: root@newsbf02.news.aol.com Message-ID: <3q367i$bg6@newsbf02.news.aol.com> Reply-To: microplate@aol.com (MICROPLATE) NNTP-Posting-Host: newsbf02.mail.aol.com I am interested in writing the history of Microtiter plates, the word I be;lieve is the trade mark of Dynatech( I do not work or represent them) however various other claim they invented the Microtiter plate also know as Microplate or cell culture or cluster dishes. Please tell me dates, who when and where. Will send you a copy of my publication for your contribution. Thanks . MICROPLATE Innovator From owner-7tms_r@net.bio.net Sun May 28 23:00:00 1995 Path: biosci!bloom-beacon.mit.edu!usc!howland.reston.ans.net!pipex!sunsite.doc.ic.ac.uk!daresbury!not-for-mail From: mcroos@imb-jena.de (Martin Roos) Newsgroups: bionet.molbio.proteins.7tms_r Subject: help: receptor binding studies Date: 29 May 1995 08:47:44 +0100 Lines: 35 Sender: lpddist@mserv1.dl.ac.uk Distribution: bionet Message-ID: <3qbu70$ial@mserv1.dl.ac.uk> Original-To: 7tms_r@dl.ac.uk Dear Netters, I am starting to establish a receptor assay, involving radioactive marked peptide hormone, in order to get both receptor number and K. I am quite familiar with the basic parts of the theory (Scatchard etc.) but still seeking for a kind of handy practical guide, which also enables one to extract non-specific binding of ligand. Any suggestions? Thanks Martin _______________________________________________________ ____________________ Martin Roos.................................. e-mail: mcroos@imb-jena.de Institut für Molekulare Biotechnologie e.V... fax: ++49-3641-656431 Proteinlabor ................................ tel: ++49-3641-656263 Beutenbergstr. 11..................................................... ...... D-07745 Jena................................................... ............. Germany _______________________________________________________________________________ From owner-7tms_r@net.bio.net Mon May 29 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!usenet.ins.cwru.edu!usenet From: Bryan Roth Newsgroups: bionet.molbio.proteins.7tms_r Subject: 5-HT receptor position Date: 30 May 1995 21:23:24 GMT Organization: Case Western Reserve University, Cleveland, Ohio (USA) Lines: 15 Message-ID: <3qg2cc$3ql@usenet.INS.CWRU.Edu> NNTP-Posting-Host: meds20785.meds.cwru.edu n NIH funded 2 year post-doctoral position is available starting September 1, 1995 in the in the lab of Bryan Roth in the Departments of Biochemistry and Psychiatry at Case Western Reserve University School of Medicine. The duties include construction and characterization of mutant serotonin receptors. Qualifications include expertise in PCR, ds-DNA sequencing and eukaryotic cell transfection. Salary is negotiable. Position is firm for 2 years with possible extension for an additional year. Further information is available by sending an e-mail request. Please reply to: roth@biocserver.cwru.edu From owner-7tms_r@net.bio.net Mon May 29 23:00:00 1995 Path: biosci!daresbury!sunsite.doc.ic.ac.uk!dundee.ac.uk!usenet From: a.mehta@dundee.ac.uk (Dr. Anil Mehta) Newsgroups: bionet.molbio.proteins.7tms_r Subject: Anti rho g protein antibody Date: 30 May 1995 16:18:41 GMT Organization: University of Dundee Lines: 4 Message-ID: <3qfgh1$3bq@dux.dundee.ac.uk> NNTP-Posting-Host: cf1.medschool.dundee.ac.uk X-Newsreader: WinVN 0.92.1 I need a clean antibody capable of detecting and immunoprecipitating this G protein in lung membranes. Anyone got any ideas about a good reliable source? Anil Mehta, internet novice From owner-7tms_r@net.bio.net Mon May 29 23:00:00 1995 Path: biosci!galaxy.ucr.edu!ratatosk.yggdrasil.com!nntp-sc.barrnet.net!nntp-hub2.barrnet.net!news.Stanford.EDU!agate!dog.ee.lbl.gov!overload.lbl.gov!emf.emf.net!news.uoregon.edu!usenet.eel.ufl.edu!news.gmi.edu!srvr1.flint.umich.edu!newsxfer.itd.umich.edu!news.itd.umich.edu!usenet From: Rick Neubig Newsgroups: bionet.molbio.proteins.7tms_r Subject: Re: help: receptor binding studies Date: 30 May 1995 10:51:43 GMT Organization: University of Michigan Lines: 37 Message-ID: <3qetbv$cl8@lastactionhero.rs.itd.umich.edu> References: <3qbu70$ial@mserv1.dl.ac.uk> NNTP-Posting-Host: warbler.med.umich.edu Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1b3 (Windows; I; 16bit) mcroos@imb-jena.de (Martin Roos) wrote: > >Dear Netters, > >I am starting to establish a receptor assay, >involving radioactive marked peptide hormone, in >order to get both receptor number and K. I am quite >familiar with the basic parts of the theory >(Scatchard etc.) but still seeking for a kind of >handy practical guide, which also enables one to >extract non-specific binding of ligand. >Any suggestions? > Martin, I'd probably start with a competition curve with tracer hot and variable cold to get an IC50. A concentration of cold 100x over the IC50 should give you a good definition of non-specific provided the plateau of the comp curve is very flat (meaning that there is no displacement of a second component). If you then do a saturation curve with variable hot you should make sure that the cold is 100x greater than both hot and Kd (if possible). As usual, pay attention to the fraction of hot bound (it should be small e.g. <10%), make sure you are really at equilibrium with a time course, and check ligand after incubation with your prep to make sure that you aren't degrading ligand under your assay conditions. Good luck! 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 May 31 23:00:00 1995 Path: biosci!bath.ac.uk!bsscw From: bsscw@bath.ac.uk (C Wood) Newsgroups: bionet.molbio.proteins.7tms_r Subject: help Date: 1 Jun 1995 03:33:49 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 21 Sender: daemon@net.bio.net Distribution: world Message-ID: Hi, By any chance does any one know the current location of Huber et al (1988). Their structure of heavy riboflavin synthase appears to me to be a noc (no co-ordinates) entry on Brookhaven (pdb0hrs.noc). I am trying to contact remnants of the group to hopefully get hold of the structure to assess its value for modelling the transmembrane regions of a receptor protein. Its use in academic research project will of course be properly acknowledged. Huber et al (1988): Ladenstein Rudolf, Schneider, M., Huber, R., Bartunik, H-D., Wilson, K., Schott, K. and Bacher, A. (1988). Heavy Riboflavin Synthase From Bacillus subtilus - Crystal Structure Analysis Of The Icoshedral B60 Capsid at 3.3 A Resolution. J. Mol. Biol. Vol 203, No 4, 1045-1070. From owner-7tms_r@net.bio.net Wed May 31 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!swrinde!gatech!bloom-beacon.mit.edu!world!decwrl!tribune.usask.ca!rover.ucs.ualberta.ca!news.ucalgary.ca!news From: Beavis N Butthead Newsgroups: bionet.biophysics,bionet.cellbiol,bionet.general,bionet.glycosci,bionet.microbiology,bionet.molbio.genbank,bionet.molbio.gene-linkage,bionet.molbio.gene-org,bionet.molbio.proteins,bionet.molbio.proteins.7tms_r Subject: Promiscuity in Protein DNA Interactions Date: 2 Jun 1995 00:22:04 GMT Organization: Terminally Unemployable Lines: 22 Message-ID: <3qlljc$qjg@ds2.acs.ucalgary.ca> NNTP-Posting-Host: acs5.acs.ucalgary.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Xref: biosci bionet.biophysics:958 bionet.cellbiol:2356 bionet.general:15527 bionet.glycosci:228 bionet.microbiology:2393 bionet.molbio.genbank:2035 bionet.molbio.gene-linkage:709 bionet.molbio.proteins:4703 bionet.molbio.proteins.7tms_r:278 The following is a description of protein-DNA interactions by a novel proto-oncogene, Dck: Dck is a transcription factor, activated by the Tyr kinase, Lck. Dck is a novel TC factor, in that it has both zinc finger and leucine zipper motifs. Upon stimulation by Lck, two Dck polypeptides homodimerizers (Not that there is anything wrong with it) to form a complex with two globular domains, with one protruding zinc finger. The Zn finger is also unusual for the critical amino acid residue for DNA interaction is His, rather than Cys. Generally, its interaction with DNA is promiscuous, but can be a sequence specific as well. The consensus sequence is CATXXXCAT. The homodimer initially binds DNA, causing a slight bend in the zinc finger motif. It then slides along the DNA in search of the CAT sequence. It will then insert at the consensus sequence at both groove majora and minora. The CAT-DNA then unwinds, allowing full insertion of the zinc finger motif. This interaction results in a kink in the DNA, relieving the torque and releasing a tremendous amount of free energy. From owner-7tms_r@net.bio.net Wed May 31 23:00:00 1995 Path: biosci!bcm!cs.utexas.edu!howland.reston.ans.net!math.ohio-state.edu!uwm.edu!newsspool.doit.wisc.edu!decwrl!tribune.usask.ca!rover.ucs.ualberta.ca!news.ucalgary.ca!news From: Beavis N Butthead Newsgroups: bionet.biophysics,bionet.cellbiol,bionet.general,bionet.glycosci,bionet.microbiology,bionet.molbio.proteins,bionet.molbio.proteins.7tms_r Subject: Promiscuity in Protein DNA Interactions Date: 2 Jun 1995 00:29:54 GMT Organization: Terminally Unemployable Lines: 21 Message-ID: <3qlm22$qkm@ds2.acs.ucalgary.ca> NNTP-Posting-Host: acs5.acs.ucalgary.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 1.1N (Windows; I; 16bit) Xref: biosci bionet.biophysics:959 bionet.cellbiol:2357 bionet.general:15528 bionet.glycosci:229 bionet.microbiology:2394 bionet.molbio.proteins:4704 bionet.molbio.proteins.7tms_r:279 The following is a description of protein-DNA interactions by a novel proto-oncogene, Dck: Dck is a transcription factor, activated by the Tyr kinase, Lck. Dck is a novel TC factor, in that it has both zinc finger and leucine zipper motifs. Upon stimulation by Lck, two Dck polypeptides homodimerizers (Not that there is anything wrong with it) to form a complex with two globular domains, with one protruding zinc finger. The Zn finger is also unusual for the critical amino acid residue for DNA interaction is His, rather than Cys. Generally, its interaction with DNA is promiscuous, but can be a sequence specific as well. The consensus sequence is CATXXXCAT. The homodimer initially binds DNA, causing a slight bend in the zinc finger motif. It then slides along the DNA in search of the CAT sequence. It will then insert at the consensus sequence at both groove majora and minora. The CAT-DNA then unwinds, allowing full insertion of the zinc finger motif. This interaction results in a kink in the DNA, relieving the torque and releasing a tremendous amount of free energy. Beavis N Butthead