From owner-proteins@hgmp.mrc.ac.uk Thu Jun 1 13:43:01 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 9EDD417ABA; Thu, 1 Jun 2000 13:43:00 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7A84B17A56; Thu, 1 Jun 2000 13:42:58 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A5A67415D4 for ; Thu, 1 Jun 2000 13:42:56 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA15527 for proteins-list@hgmp.mrc.ac.uk; Thu, 1 Jun 2000 13:42:55 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Christina Benedek" X-Newsgroups: bionet.molbio.proteins Subject: protein synthesis - question Date: Thu, 1 Jun 2000 14:42:05 +0200 Lines: 23 Message-ID: <8h5loc$2d2je$2@fu-berlin.de> X-Trace: fu-berlin.de 959863373 2525806 149.225.91.231 (16 [25865]) X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2014.211 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2014.211 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hello! I hope I'm right in this ng. If not: could you please tell me, which ng would be right for tis question? Thanks I'm preparing for a test I have got a question regarding plasma membrane proteins and can't find the answer in any of the books I got at home and don't know where to search on th web for the answer. So I hope you can help me. Plasma membrane proteins are synthesised with the help of ribosomes attached to the ER, they are simultaneously translocated into the lumen of the ER. But what then? They need to be integrated into the ER membrane before being transported elsewhere. But how? I can imagine this is as difficult as a protein flip-flop in the membrane would be. Does anyone know the answer? Thanks for reply C. Benedek From owner-proteins@hgmp.mrc.ac.uk Thu Jun 1 17:40:16 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id CE67B17B4F; Thu, 1 Jun 2000 17:40:15 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id C859F17AD5; Thu, 1 Jun 2000 17:40:13 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 30F2B415CA for ; Thu, 1 Jun 2000 17:40:13 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA18354 for proteins-list@hgmp.mrc.ac.uk; Thu, 1 Jun 2000 17:40:12 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: strider@comteco.entelnet.bo X-Newsgroups: bionet.molbio.proteins Subject: Re: MALDI-software Date: Thu, 01 Jun 2000 16:29:52 GMT Organization: Deja.com - Before you buy. Lines: 14 Message-ID: <8h631j$4pr$1@nnrp1.deja.com> References: <3919C4C5.6591A722@ruhr-uni-bochum.de> <8fq1kb$c1b$1@sunb.ocs.mq.edu.au> X-Article-Creation-Date: Thu Jun 01 16:29:52 2000 GMT X-Http-User-Agent: Mozilla/4.72 [en] (X11; U; Linux 2.2.14-5.0 i686) X-Http-Proxy: 1.0 x67.deja.com:80 (Squid/1.1.22) for client 166.114.106.164 X-MyDeja-Info: XMYDJUIDdavor To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hello Tatjana, No news group but you can take a look at www.expasy.ch or one of its mirrors.... lots of related software.... you might want to try expasy.ucbcba.edu.bo (we are in a test phase and will soon be a mirror.... -> less traffic davor Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Thu Jun 1 19:33:46 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7029317ACE; Thu, 1 Jun 2000 19:33:45 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 25E4217ABB; Thu, 1 Jun 2000 19:33:44 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 96E74415CA for ; Thu, 1 Jun 2000 19:33:43 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA19819 for proteins-list@hgmp.mrc.ac.uk; Thu, 1 Jun 2000 19:33:42 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Michael Hofle X-Newsgroups: bionet.molbio.proteins Subject: nucleotide sequences for PDB proteins???.. Date: Thu, 01 Jun 2000 13:24:19 -0700 Organization: Texas A&M University, College Station, Texas Lines: 7 Message-ID: <3936C673.597B7F8@tamu.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.6 [en] (WinNT; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Does anyone know where I could find the nucleotide sequences for proteins listed in the PDB. The PDB accession numbers don't cross list with the Genbank numbers. Any help would be greatly appreciated!! Thanks, Michael From owner-proteins@hgmp.mrc.ac.uk Fri Jun 2 09:25:09 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7AF5F17AA7; Fri, 2 Jun 2000 09:25:08 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D3B0D17AA1; Fri, 2 Jun 2000 09:25:06 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 1219E415DF for ; Fri, 2 Jun 2000 09:25:06 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id JAA08035 for proteins-list@hgmp.mrc.ac.uk; Fri, 2 Jun 2000 09:25:05 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Cornelius Krasel Subject: Re: protein synthesis - question X-Newsgroups: bionet.molbio.proteins References: <8h5loc$2d2je$2@fu-berlin.de> Organization: =?ISO-8859-1?Q?Universit=E4t_W=FCrzburg?= User-Agent: tin/pre-1.4-19990805 ("Preacher Man") (UNIX) (Linux/2.2.13 (i686)) Date: Fri, 2 Jun 2000 10:16:06 +0200 Message-ID: <6gq7h8.e0d.ln@wpxx02.toxi.uni-wuerzburg.de> X-Trace: 2 Jun 2000 10:25:02 +0100, wpxx02.toxi.uni-wuerzburg.de Lines: 20 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Christina Benedek wrote: > Plasma membrane proteins are synthesised with the help of ribosomes attached > to the ER, they are simultaneously translocated into the lumen of the ER. No, they are integrated into the ER membrane in the correct topology during synthesis. The exact mechanism is still unknown (especially for proteins which contain more than one transmembrane domain). A rough overview should be contained in about any recent biochemistry or cell biology book. Don't be afraid of the English versions; most of the books are usually written in a very clear style which (IMO) often gets lost in the translation to German. You will also need the English in your future career anyway. HTH, --Cornelius. -- /* Cornelius Krasel, U Wuerzburg, Dept. of Pharmacology, Versbacher Str. 9 */ /* D-97078 Wuerzburg, Germany email: phak004@rzbox.uni-wuerzburg.de SP4 */ /* "Science is the game we play with God to find out what His rules are." */ From owner-proteins@hgmp.mrc.ac.uk Fri Jun 2 14:37:26 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id DD91817B93; Fri, 2 Jun 2000 14:37:25 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7984717B8C; Fri, 2 Jun 2000 14:37:24 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id B2E0C415CA for ; Fri, 2 Jun 2000 14:37:23 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA11790 for proteins-list@hgmp.mrc.ac.uk; Fri, 2 Jun 2000 14:37:23 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: fedmahn@ihug.com.au X-Newsgroups: bionet.molbio.proteins Subject: Nuclear Extract Date: Fri, 02 Jun 2000 23:35:07 +1000 Organization: The Internet Group (Sydney) Lines: 10 Message-ID: <3937B80B.47718BCF@ihug.com.au> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Server-Date: 2 Jun 2000 13:36:14 GMT X-Mailer: Mozilla 4.72 [en] (Win98; U) X-Accept-Language: zh-TW,en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, I am trying to get nuclear extracts from whole organs (rats or sheep), does anyone know of any fast and non-detergent method? And any idea on good nuclear, cytoplasmic and mitochondrial protein markers? (I get a lot of contaminants in my previous preps, I want to be sure that the nuclear extract is CLEAN). Many thanks in advance. From owner-proteins@hgmp.mrc.ac.uk Fri Jun 2 20:45:59 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id CD88317B2F; Fri, 2 Jun 2000 20:45:58 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id A277617B2D; Fri, 2 Jun 2000 20:45:57 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 5760D415DA for ; Fri, 2 Jun 2000 20:45:56 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA16338 for proteins-list@hgmp.mrc.ac.uk; Fri, 2 Jun 2000 20:45:55 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: karthi@uic.edu ("Dr. Karthikeyan Narayanan") X-Newsgroups: bionet.molbio.proteins Subject: mass analysis (ESI) Date: 2 Jun 2000 20:45:54 +0100 Organization: UIC Lines: 20 Message-ID: <39380FF6.F13A6C32@uic.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 959975155 16336 193.62.192.80 (2 Jun 2000 19:45:55 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D01E317B2F; Fri, 2 Jun 2000 20:45:49 +0100 (BST) X-Received: from eeyore.cc.uic.edu (eeyore.cc.uic.edu [128.248.171.51]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 773BE415DA for ; Fri, 2 Jun 2000 20:45:48 +0100 (BST) X-Received: from uic.edu (den-500c.hub.uic.edu [128.248.79.50]) by eeyore.cc.uic.edu (8.9.3/8.9.3) with ESMTP id OAA09742 for ; Fri, 2 Jun 2000 14:45:47 -0500 (CDT) X-Reply-To: karthi@uic.edu X-Mailer: Mozilla 4.72 [en] (Win98; I) X-Accept-Language: en X-To: proteins@net.bio.net To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, I want to analyze an extremely acidic protein using ESI mass spectrometry. However, I could not get any apparent signal. could any one give me any suggestions? -- ******************************************************************************************** Narayanan Karthikeyan Ph.D., College of Dentistry, Department of Oral Biology, University of Illinois at Chicago Chicago IL 60612 Tel: 312-413-5375 Fax: 312-413-6694 ******************************************************************************************** --- From owner-proteins@hgmp.mrc.ac.uk Mon Jun 5 09:14:36 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3B9C117B01; Mon, 5 Jun 2000 09:14:35 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 9024817AFC; Mon, 5 Jun 2000 09:14:33 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D9F16415D4 for ; Mon, 5 Jun 2000 09:14:32 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id JAA24590 for proteins-list@hgmp.mrc.ac.uk; Mon, 5 Jun 2000 09:14:32 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Frank Fuerst X-Newsgroups: bionet.molbio.proteins Subject: Re: nucleotide sequences for PDB proteins???.. Date: Mon, 05 Jun 2000 10:16:03 +0200 Lines: 14 Message-ID: <8hfnh6$2vmve$1@fu-berlin.de> References: <3936C673.597B7F8@tamu.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: fu-berlin.de 960192870 3136494 141.89.201.37 (16 49 955) X-Mailer: Mozilla 4.51 [de]C-CCK-MCD QXW03200 (WinNT; I) X-Accept-Language: de,en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Michael Hofle wrote: > > Does anyone know where I could find the nucleotide sequences for > proteins listed in the PDB. The PDB accession numbers don't cross list > with the Genbank numbers. Any help would be greatly appreciated!! Try http://www.expasy.ch/cgi-bin/sprot-search-ful at the SwissProt Database. Frank -- Die Verwendung von mehreren Ausrufezeichen macht die Aussage nicht ausrufender sondern ausufernder. [Michael Bauer in dnq] From owner-proteins@hgmp.mrc.ac.uk Tue Jun 6 14:10:15 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id EC52B17B7F; Tue, 6 Jun 2000 14:10:14 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7A4AA17B82; Tue, 6 Jun 2000 14:10:13 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 065E2415DC for ; Tue, 6 Jun 2000 14:10:10 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA24085 for proteins-list@hgmp.mrc.ac.uk; Tue, 6 Jun 2000 14:10:10 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: strider@comteco.entelnet.bo X-Newsgroups: bionet.molbio.proteins Subject: need peak detection algorithms Date: Tue, 06 Jun 2000 12:57:36 GMT Organization: Deja.com - Before you buy. Lines: 23 Message-ID: <8hisfn$t4r$1@nnrp1.deja.com> X-Article-Creation-Date: Tue Jun 06 12:57:36 2000 GMT X-Http-User-Agent: Mozilla/4.72 [en] (X11; U; Linux 2.2.14-5.0 i686) X-Http-Proxy: 1.0 x55.deja.com:80 (Squid/1.1.22) for client 166.114.106.8 X-MyDeja-Info: XMYDJUIDdavor To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hello, I am working on de Novo peptide sequencing with MS/MS data. I am now trying to learn all the different ion types generated by the MS/MS so that I can assign some probabilities to each terminal ion type... say y, b, y-H2O, b-H2O, etc. I have statistically obtained plots for the ferquency functions of: - N-term charge +1 - C-term charge +1 - N-term charge +2 - C-term charge +2 What I need now is a very good peak detection algorithm that considers random fluctuations and false peaks.... Can anybody helpme on this? I mean the algorithms.... thanx Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Tue Jun 6 14:22:25 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C328C17B7C; Tue, 6 Jun 2000 14:22:24 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id BF2C517B71; Tue, 6 Jun 2000 14:22:22 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 1A916415D4 for ; Tue, 6 Jun 2000 14:22:22 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA24225 for proteins-list@hgmp.mrc.ac.uk; Tue, 6 Jun 2000 14:22:21 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: brookes@uab.edu ("Paul S. Brookes.") X-Newsgroups: bionet.molbio.proteins Subject: Re: Nuclear Extract Date: 6 Jun 2000 14:22:20 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 63 Message-ID: <4.3.1.2.20000606081752.00ade3d0@norm.dpo.uab.edu> Mime-Version: 1.0 Content-Type: multipart/alternative; boundary="=====================_59139718==_.ALT" X-Trace: niobium.hgmp.mrc.ac.uk 960297741 24223 193.62.192.80 (6 Jun 2000 13:22:21 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 3C88B17B7C; Tue, 6 Jun 2000 14:22:15 +0100 (BST) X-Received: from lilith2.dpo.uab.edu (lilith.dpo.uab.edu [138.26.1.128]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id AF3E7415D5 for ; Tue, 6 Jun 2000 14:22:12 +0100 (BST) X-Received: from pathvhg004c.uab.edu ([138.26.154.15]) by lilith2.dpo.uab.edu (8.9.3/8.9.3) with ESMTP id IAA03457 for ; Tue, 6 Jun 2000 08:22:11 -0500 X-Sender: brookes@norm.dpo.uab.edu X-Mailer: QUALCOMM Windows Eudora Version 4.3.1 X-To: proteins@net.bio.net To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk --=====================_59139718==_.ALT Content-Type: text/plain; charset="us-ascii"; format=flowed By nuclear extracts do you mean just the DNA (in which case a simple precipitation method should work) or the whole nucleus including the membrane and any proteins there - which is a bit more tricky? WRT markers, LDH is about as good a cytosolic marker as any - rock solid spectrophotometric rates and lots of kits available from Sigma e.t.c. if you can't be bothered to work up a method. Mitochondrial markers are a bit more tricky - try citrate synthase (a matrix enzyme), as its easy to assay. However your contamination might be from the mito' membrane fraction, so try monoamine oxidase (outer membrane) or cytochrome c oxidase (inner membrane). PSB _________________________________________ Dr. Paul S. Brookes. (brookes@uab.edu) UAB Department of Pathology, G004 Volker Hall 1670 University Blvd., Birmingham AL 35294 USA Tel (001) 205 934 1915 Fax (001) 205 934 1775 http://peir.path.uab.edu/brookes The quality of e-mails can go down as well as up --=====================_59139718==_.ALT Content-Type: text/html; charset="us-ascii"
By nuclear extracts do you mean just the DNA (in which case a simple precipitation method should work) or the whole nucleus including the membrane and any proteins there - which is a bit more tricky?

WRT markers, LDH is about as good a cytosolic marker as any - rock solid spectrophotometric rates and lots of kits available from Sigma e.t.c. if you can't be bothered to work up a method.  Mitochondrial markers are a bit more tricky - try citrate synthase (a matrix enzyme), as its easy to assay.  However your contamination might be from the mito' membrane fraction, so try monoamine oxidase (outer membrane) or cytochrome c oxidase (inner membrane).

PSB

_________________________________________
Dr. Paul S. Brookes.            (brookes@uab.edu)
UAB Department of Pathology,   G004 Volker Hall
1670 University Blvd., Birmingham AL 35294 USA
Tel (001) 205 934 1915     Fax (001) 205 934 1775
http://peir.path.uab.edu/brookes

The quality of e-mails can go down as well as up
 --=====================_59139718==_.ALT-- --- From owner-proteins@hgmp.mrc.ac.uk Wed Jun 7 13:58:41 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C6A6C17B6F; Wed, 7 Jun 2000 13:58:40 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 9DE7517B6E; Wed, 7 Jun 2000 13:58:39 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 21E1F415CF for ; Wed, 7 Jun 2000 13:58:39 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA25326 for proteins-list@hgmp.mrc.ac.uk; Wed, 7 Jun 2000 13:58:38 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f Message-ID: <393E46F5.CE55EA4D@uni-konstanz.de> Date: Wed, 07 Jun 2000 14:58:32 +0200 From: "Frank O. Fackelmayer" Reply-To: Frank.Fackelmayer@uni-konstanz.de X-Mailer: Mozilla 4.72 (Macintosh; I; PPC) X-Accept-Language: en MIME-Version: 1.0 X-Newsgroups: bionet.molbio.proteins Subject: Re: blue native gel electrophoresis References: <200005241942.AA01192@mani.cbs.umn.edu> <1ebacnp.chbu6v14rfow0N@stol-117-182.uva.studentennet.nl> Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Trace: 7 Jun 2000 14:58:29 +0100, 134.34.126.40 Organization: University of Constance, Germany Lines: 31 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Pathos wrote: > > In article <1ebacnp.chbu6v14rfow0N@stol-117-182.uva.studentennet.nl>, > jacqg@zpam.dds.nl (jw) wrote: > > > look for papers by schagger (umlaut on the a) > > When writting umlauts in german without the umlauts, simple put an "e" > after the vowel that needs an umlaut. So it would be written von > Schaegger. > > I see that your address is from the .nl region of the world so I suspect > you already knew that. > > regards, > Peter Pediaditakis > Well, yes Peter, you are right. That is exactly the way we germans write it when there are no umlaut characters available (the german alphabet has three umlauts, the vowels a,o, and u with two dots on top; they are pronounced very differently from the vowels without dots; e.g. "a umlaut" is pronounced quite like "ai" in "pair", whereas the plain "a" is like "a" in "father"). The problem is that Medline is an all-american thing that does not care about umlauts in names. Thus, any "a umlaut" (ä) will become a simple "a" in medline entries. It is up to the authors to be aware of that and write their own names without umlaut characters. Some do, others dont. So, it is safer to search for e.g. ("SCHAGGER" OR "SCHAEGGER"). Frank From owner-proteins@hgmp.mrc.ac.uk Wed Jun 7 17:29:34 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 2295317B6F; Wed, 7 Jun 2000 17:29:34 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D5E5517B5F; Wed, 7 Jun 2000 17:29:32 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E1CAE415D5 for ; Wed, 7 Jun 2000 17:29:31 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA28806 for proteins-list@hgmp.mrc.ac.uk; Wed, 7 Jun 2000 17:29:31 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Arlin Stoltzfus X-Newsgroups: bionet.molbio.proteins Subject: Re: nucleotide sequences for PDB proteins???.. Date: Wed, 07 Jun 2000 12:24:38 -0400 Organization: CARB Lines: 21 Message-ID: <393E7746.30E4192F@carb.nist.gov> References: <3936C673.597B7F8@tamu.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: hecate.umd.edu 960395370 31999 129.6.113.43 (7 Jun 2000 16:29:30 GMT) X-Complaints-To: abuse@wam.umd.edu X-Mailer: Mozilla 4.73 [en] (X11; U; Linux 2.2.14-6.0.1smp i686) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Michael Hofle wrote: > > Does anyone know where I could find the nucleotide sequences for > proteins listed in the PDB. The PDB accession numbers don't cross list > with the Genbank numbers. Any help would be greatly appreciated!! Every PDB structure is in the MMDB, NCBI's value-added molecular structure database, and every structure in the MMDB is linked to an amino acid sequence in the protein subdivision of GenBank. If there is a gene entry corresponding to the protein, there will be a link to this as well (though its not guaranteed that the gene has been sequenced for every PDB chain). You can get all of this from the Entrez web interface just by starting with a structure entry and then using the pull-down menu to select "protein links" or "nucleotide links". Arlin -- Arlin Stoltzfus (arlin@carb.nist.gov) CARB, 9600 Gudelsky Drive, Rockville, Md 20850 ph. 301 738-6208; fax 301 738-6255 From owner-proteins@hgmp.mrc.ac.uk Wed Jun 7 17:42:25 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id A34F817B73; Wed, 7 Jun 2000 17:42:24 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 57B8717B72; Wed, 7 Jun 2000 17:42:23 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id AC420415D4 for ; Wed, 7 Jun 2000 17:42:22 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA29042 for proteins-list@hgmp.mrc.ac.uk; Wed, 7 Jun 2000 17:42:22 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: hooverd@ncifcrf.gov ("Dr. David Hoover") X-Newsgroups: bionet.molbio.proteins Subject: synthetic gene software Date: 7 Jun 2000 17:42:21 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 15 Message-ID: <4.1.20000607123648.00a4d100@mail.ncifcrf.gov> Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" X-Trace: niobium.hgmp.mrc.ac.uk 960396141 29040 193.62.192.80 (7 Jun 2000 16:42:21 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 2F89417B72; Wed, 7 Jun 2000 17:42:19 +0100 (BST) X-Received: from mail.ncifcrf.gov (mail.NCIFCRF.GOV [129.43.100.100]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 6CDD1415D4 for ; Wed, 7 Jun 2000 17:42:18 +0100 (BST) X-Received: from crypc116 ([129.43.17.36]) by mail.ncifcrf.gov (8.10.1/8.10.1) with SMTP id e57GgLL14522 for ; Wed, 7 Jun 2000 12:42:21 -0400 (EDT) X-Sender: hooverd@mail.ncifcrf.gov X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1 X-To: proteins@hgmp.mrc.ac.uk To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Does anyone know of shareware that will take an amino acid sequence as input, generates a nucleotide sequence optimized for expression in E. coli and restriction sites and outputs oligonucleotides (potentially hundreds) for generating a synthetic gene? Thanks, David Hoover ======================================== Dr. David Hoover, Postdoctoral Fellow NCI-FCRDC, Macromolecular Structure Laboratory Bldg. 539, Ft. Detrick MD 21702 PH: 301-846-5326, FAX: 301-846-7101 --- From owner-proteins@hgmp.mrc.ac.uk Thu Jun 8 11:20:07 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5B45217AB0; Thu, 8 Jun 2000 11:20:06 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D86FD17AAD; Thu, 8 Jun 2000 11:20:04 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 6175A415CF for ; Thu, 8 Jun 2000 11:20:04 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id LAA20171 for proteins-list@hgmp.mrc.ac.uk; Thu, 8 Jun 2000 11:20:04 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Dr. Duncan Clark" X-Newsgroups: bionet.molbio.proteins Subject: Liposomes Date: Thu, 8 Jun 2000 11:13:32 +0100 Organization: DNAmp Ltd. Message-ID: Reply-To: "Dr. Duncan Clark" X-NNTP-Posting-Host: genesys.demon.co.uk:158.152.17.110 X-Trace: news.demon.co.uk 960459592 nnrp-14:18778 NO-IDENT genesys.demon.co.uk:158.152.17.110 X-Complaints-To: abuse@demon.net MIME-Version: 1.0 X-Newsreader: Turnpike Integrated Version 5.01 M Lines: 34 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi Folks, I'm kind of stuck and doing a general web search with 'liposomes' as the search word gives thousands of hits with little info. What I want to know, as I know very little about liposomes, is: Is there any good web site on liposomes in general? Can one incorporate proteins in liposomes? Is it easy to do and if so can anyone point me to a reference or web site. How stable are liposomes in terms of temperature? Am I right in assuming they 'melt' or fall apart and if so what is the maximum temp they can be used at (again I assume this is somewhat dependant upon the type of reagents using to construct them) ? I am way out of my own field here, so any pointers really appreciated. Many thanks Duncan -- The problem with being on the cutting edge is that you occasionally get sliced from time to time.... Duncan Clark GeneSys Ltd. Tel: +44(0)1252376288 FAX: +44(0)8701640382 From owner-proteins@hgmp.mrc.ac.uk Thu Jun 8 19:30:06 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1C9EC17AA5; Thu, 8 Jun 2000 19:30:06 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id CD4A817A59; Thu, 8 Jun 2000 19:30:04 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 54A06415D4 for ; Thu, 8 Jun 2000 19:30:04 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA26339 for proteins-list@hgmp.mrc.ac.uk; Thu, 8 Jun 2000 19:30:03 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Chris Wiethoff X-Newsgroups: bionet.molbio.proteins Subject: Re: Liposomes Date: Thu, 8 Jun 2000 13:29:44 -0500 Organization: University of Kansas Computing Services Lines: 26 Message-ID: References: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII In-Reply-To: To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk On Thu, 8 Jun 2000, Dr. Duncan Clark wrote: > Hi Folks, > > I'm kind of stuck and doing a general web search with 'liposomes' as the > search word gives thousands of hits with little info. > > What I want to know, as I know very little about liposomes, is: > > Is there any good web site on liposomes in general? http://members.tripod.com/~rkrishna/ The above website is full of tons of information regarding liposomes including reference citations for various applications such as the incorporation of proteins into liposomes. Additionally, you should be able to find answers regarding your 'stability' related questions on this site. Hope this helps, Chris From owner-proteins@hgmp.mrc.ac.uk Thu Jun 8 20:45:23 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C688817AA5; Thu, 8 Jun 2000 20:45:21 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 4205A17AA1; Thu, 8 Jun 2000 20:45:20 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 9F9A0415D4 for ; Thu, 8 Jun 2000 20:45:19 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA27129 for proteins-list@hgmp.mrc.ac.uk; Thu, 8 Jun 2000 20:45:19 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "dawe" X-Newsgroups: bionet.molbio.proteins Subject: Ab and phaseolin Lines: 8 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 Message-ID: Date: Thu, 08 Jun 2000 19:44:35 GMT X-Complaints-To: abuse@libero.it X-Trace: news.infostrada.it 960493475 151.20.39.221 (Thu, 08 Jun 2000 21:44:35 MET DST) Organization: [Infostrada] To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In a western blotting of total protein extraction by tobacco and marked with antibodies anti BiP and anti phaseolin I've found a band of approx. 32-34 kDa (I've found the BiP band too...). Can anybody give me an idea of what that band is? Davide From owner-proteins@hgmp.mrc.ac.uk Fri Jun 9 19:37:50 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id E833517B4F; Fri, 9 Jun 2000 19:37:49 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 5CD9817B4C; Fri, 9 Jun 2000 19:37:48 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id B6CAB415D5 for ; Fri, 9 Jun 2000 19:37:47 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA22730 for proteins-list@hgmp.mrc.ac.uk; Fri, 9 Jun 2000 19:37:47 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Deborah_Britt@brown.edu (Deb Britt) X-Newsgroups: bionet.molbio.proteins Subject: FLAG epitope and protein migration on PAGE Date: Fri, 09 Jun 2000 14:36:38 -0400 Organization: Rhode Island Hospital Lines: 28 Message-ID: X-Newsreader: MT-NewsWatcher 2.3.3b4 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I was wondering if anyone has ever noticed anomalous migration during PAGE of proteins tagged with the FLAG epitope. We used a monoclonal antibody to screen a library and obtain a cDNA for the corresponding antigen. In Western blots, the monoclonal detects a 69 kD band in normal rat liver extract, and so does an antipeptide antibody raised to the protein predicted from the cloned cDNA. However, we put the cDNA in an expression vector so the protein is expressed with the FLAG epitope fused to it, and when we run the Western the band is at 97 kD, as detected by anti-FLAG, the monoclonal and the anti-peptide abs. I am at a loss to explain the size difference. According to sequence data, the start and stop codons are ok, and the size predictions by computer analysis say 65 kd, a reasonable match to the 69 kD we see in the native protein. We are in the process of trying to cleave off the FLAG prior to running the gel, and we're also putting the protein in a different expression system, but I was curious to find out whether anyone else has ever had a similar experience, or can offer any suggestions as to what might be happening. Thanks. -- Deborah Britt, Ph.D. Department of Medical Oncology Rhode Island Hospital From owner-proteins@hgmp.mrc.ac.uk Sat Jun 10 23:11:56 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 540C717B96; Sat, 10 Jun 2000 23:11:54 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 8A3D217AB5; Sat, 10 Jun 2000 23:11:49 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 0B806415DD for ; Sat, 10 Jun 2000 23:11:49 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id XAA00159 for proteins-list@hgmp.mrc.ac.uk; Sat, 10 Jun 2000 23:11:48 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: gwh@spam.block.cpcug.org (Glen Humphrey) X-Newsgroups: bionet.molbio.proteins Subject: Re: FLAG epitope and protein migration on PAGE Message-ID: <3942b7f8.4832752@news.randori.com> References: X-Newsreader: Forte Free Agent 1.11/32.235 Lines: 22 X-Trace: news.randori.com 960675059 (Sat, 10 Jun 2000 15:10:59 PDT) Organization: Randori News -- http://www.randori.com X-Sender: agent86 Date: Sat, 10 Jun 2000 22:11:02 GMT To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk On Fri, 09 Jun 2000 14:36:38 -0400, Deborah_Britt@brown.edu (Deb Britt) wrote: >I was wondering if anyone has ever noticed anomalous migration during PAGE >of proteins tagged with the FLAG epitope. > Yes, adding the Flag epitope to a protein will often cause it to migrate a little slower in SDS-PAGE. This is partly due to the additional mass and partly due to the Flag sequence being rather hydrophillic, such that it binds relatively less SDS than a "typical" protein sequence of the same length. Other hydrophillic modifications to proteins have the same effect, i.e. phosphorylation. The reduced mobility caused by the Flag epitope can be an advantage, if you want to distinguish the Flag-tagged recombinant protein from its endogenous counterpart (i.e. expressed in transfected cells). - Glen -> To reply, delete "spam.block." from my email address. From owner-proteins@hgmp.mrc.ac.uk Mon Jun 12 16:03:17 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 41E3817AD5; Mon, 12 Jun 2000 16:03:16 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id DA99F17A79; Mon, 12 Jun 2000 16:03:13 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 66344415DA for ; Mon, 12 Jun 2000 16:03:13 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA07312 for proteins-list@hgmp.mrc.ac.uk; Mon, 12 Jun 2000 16:03:12 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: hart@chinacat.uthscsa.edu (P. John Hart) X-Newsgroups: bionet.molbio.proteins Subject: Postdoctoral Positions in Crystallography Date: 12 Jun 2000 14:54:31 GMT Organization: UT Health Science Center, San Antonio Lines: 31 Distribution: world Message-ID: <8i2tj7$t9o$3@cosmos.uthscsa.edu> Reply-To: pjhart@biochem.uthscsa.edu (P. John Hart) X-Newsreader: mxrn 6.18-10 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk POSTDOCTORAL POSITIONS X-RAY CRYSTALLOGRAPHY Two funded POSTDOCTORAL POSITIONS are available immediately to examine structure/function relationships of medically relevant protein molecules using X-ray crystallography and other biophysical techniques. Both positions require previous experience in macromolecular crystallography OR extensive experience in protein structure/function analysis and heterologous protein expression. Current projects include antioxidant proteins and their derivatives, copper chaperone proteins, blue copper electron transfer proteins, and platelet-activating factor (PAF) metabolizing enzymes. Salaries will range from $25,000 - $35,000 per year, commensurate with qualifications. The laboratory is part of the X-ray Crystallographic Core facility in the Center for Biomolecular Structure Analysis located in the new Allied Health Research Building. State-of-the-art crystallographic (R-Axis IV area detector, Osmic confocal optics, RU-300 generator, X-STREAM) and computational (DEC Alphas/SGIs) facilities are in place and are fully functional. Ample shared facilities (high-field NMR, protein core, DNA sequencing, mass spec, analytical ultracentrifugation, etc.) are also available in the Department of Biochemistry. Applicants should hold a Ph.D. in biochemistry, chemistry, molecular biology, or a related field. Interested persons are asked to send their curriculum vitae along with the names and addresses for three letters of reference to: Dr. P. John Hart, Center for Biomolecular Structure Analysis, Department of Biochemistry, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900. Email: pjhart@biochem.uthscsa.edu. Website: www.biochem.uthscsa.edu The University of Texas Health Science Center at San Antonio is an Equal Employment Opportunity/Affirmative Action Employer. From owner-proteins@hgmp.mrc.ac.uk Mon Jun 12 16:15:08 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C2B8117A79; Mon, 12 Jun 2000 16:15:07 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7705117A6D; Mon, 12 Jun 2000 16:15:06 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E4BFE415DA for ; Mon, 12 Jun 2000 16:15:05 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id QAA07445 for proteins-list@hgmp.mrc.ac.uk; Mon, 12 Jun 2000 16:15:05 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Dr. Duncan Clark" X-Newsgroups: bionet.molbio.proteins Subject: Re: Liposomes Date: Mon, 12 Jun 2000 11:11:46 +0100 Organization: DNAmp Ltd. Message-ID: References: Reply-To: "Dr. Duncan Clark" X-NNTP-Posting-Host: genesys.demon.co.uk:158.152.17.110 X-Trace: news.demon.co.uk 960822886 nnrp-13:12110 NO-IDENT genesys.demon.co.uk:158.152.17.110 X-Complaints-To: abuse@demon.net MIME-Version: 1.0 X-Newsreader: Turnpike Integrated Version 5.01 M Lines: 20 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk In article , Chris Wiethoff writes >> Is there any good web site on liposomes in general? > >http://members.tripod.com/~rkrishna/ Just what I wanted. Many thanks Duncan -- The problem with being on the cutting edge is that you occasionally get sliced from time to time.... Duncan Clark GeneSys Ltd. Tel: +44(0)1252376288 FAX: +44(0)8701640382 From owner-proteins@hgmp.mrc.ac.uk Tue Jun 13 17:39:11 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 09E1917B0C; Tue, 13 Jun 2000 17:39:10 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id D79AC17ABD; Tue, 13 Jun 2000 17:39:09 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 6357C415DC for ; Tue, 13 Jun 2000 17:39:09 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA04303 for proteins-list@hgmp.mrc.ac.uk; Tue, 13 Jun 2000 17:39:08 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f X-Newsgroups: bionet.molbio.proteins From: r.j.siviter@leeds.ac.uk (Richard Siviter) Subject: Postdoctoral position: zinc-metallopeptidases X-Accept-Language: en Message-ID: <39465BE9.842007AC@leeds.ac.uk> X-Mailer: Mozilla 4.61 [en] (Win95; I) Content-Type: text/plain; charset=iso-8859-1 MIME-Version: 1.0 Date: Tue, 13 Jun 2000 17:17:49 +0100 (BST) Lines: 22 Content-Transfer-Encoding: 8bit To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk University of Leeds Faculty of Biological Sciences Schools of Biology and Biochemistry and Molecular Biology Research Fellow (PhD) in Biochemistry/Molecular Biology Ref. No. 083/235/004/027 This post, funded through the Genomics in Animal Function Initiative, is available immediately for a fixed period of up to three years to study the physiological roles of novel zinc-metallopeptidase genes in the fly, Drosophila melanogaster. Applicants should have a PhD in biochemistry/molecular biology or an allied subject and need not have experience of working with Drosophila. Salary will normally be on the scale for Research Staff Grade 1A starting at £16,286 -18,185 p.a. depending on qualifications and experience. For further information, see http://www.biology.leeds.ac.uk/research/Develop/index.htm. Informal enquiries about the post may be made to Dr R E Isaac, email: r.e.isaac@leeds.ac.uk, tel. 0113 2332903. Application forms and further particulars may be obtained from Mrs Lynn Buckley-McDonald, tel. 0113 233 2880, fax 0113 233 3091, School of Biology, The University of Leeds, Leeds, LS2 9JT. In all enquiries please quote the appropriate reference number From owner-proteins@hgmp.mrc.ac.uk Wed Jun 14 05:35:41 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5860217B2E; Wed, 14 Jun 2000 05:35:40 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id CEA8117B2F; Wed, 14 Jun 2000 05:35:37 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 683B2415DE for ; Wed, 14 Jun 2000 05:35:36 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id FAA21211 for proteins-list@hgmp.mrc.ac.uk; Wed, 14 Jun 2000 05:35:35 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Roger Murphy X-Newsgroups: bionet.molbio.proteins Subject: Pierce "gentle elution buffer" Date: Wed, 14 Jun 2000 14:35:34 +1000 Organization: The University of Melbourne Lines: 14 Message-ID: <39470B96.BDF27AD8@ludwig.edu.au> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: ariel.ucs.unimelb.edu.au 960957272 25802 128.250.186.193 (14 Jun 2000 04:34:32 GMT) X-Complaints-To: news@news.unimelb.edu.au X-Mailer: Mozilla 4.7 [en] (WinNT; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk A question for the assembled throng.... We're going to use the Pierce "gentle elution buffer" to elute an acid-sensitive monoclonal antibody from a protein A column. The Pierce instructions warn of dire precipitation consequences if this buffer ever comes into contact with phosphate salts... Does anyone know why? What is the ingredient in their buffer that will precipitate with phosphates? Thanks for your anticipated help! Roger From owner-proteins@hgmp.mrc.ac.uk Wed Jun 14 23:08:59 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 304BB17AF6; Wed, 14 Jun 2000 23:08:59 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id B750B17AF4; Wed, 14 Jun 2000 23:08:57 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A18A7415DD for ; Wed, 14 Jun 2000 23:08:56 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id XAA12853 for proteins-list@hgmp.mrc.ac.uk; Wed, 14 Jun 2000 23:08:56 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: p32@rocketmail.com (che pillay) X-Newsgroups: bionet.molbio.proteins Subject: redox buffer novice Date: 14 Jun 2000 23:08:55 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 23 Message-ID: <39480278.83B239E8@rocketmail.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 961020535 12850 193.62.192.80 (14 Jun 2000 22:08:55 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 8D17617AF4; Wed, 14 Jun 2000 23:08:53 +0100 (BST) X-Received: from unpsun2.cc.unp.ac.za (Unpsun2.cc.unp.ac.za [143.128.64.3]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 871E7415DA for ; Wed, 14 Jun 2000 23:08:48 +0100 (BST) X-Received: from pc-01 (sspc081.cc.unp.ac.za [143.128.160.81]) by unpsun2.cc.unp.ac.za (8.9.3/8.9.3) with ESMTP id AAA10670 for ; Thu, 15 Jun 2000 00:02:58 +0200 (SATS) X-Mailer: Mozilla 4.01 [en] (WinNT; I) X-To: proteins@net.bio.net X-Priority: 3 (Normal) X-Corel-MessageType: EMail To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi All, I am studying the redox properties of my enzyme using various redox buffers (GSH/GSSG; cysteine:cystine; DTTred/DTToxid). Most of the protocols that have described in the literature for these buffers don't describe whether these buffers were deoxygenated prior to use; or whether any precautions were taken to prevent oxidation during the assay. Are there any protocols for redox buffers that take the effect of oxygen into account ? One more question: the active site cysteines of may enzyme may become oxidised to sulfinic or sulfenic acids ... how could I test this ? Thank you for your time, Che Pillay Dept. of Biochemsitry University of Natal South Africa pillayche@nu.ac.za --- From owner-proteins@hgmp.mrc.ac.uk Thu Jun 15 15:35:15 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C520D17ACF; Thu, 15 Jun 2000 15:35:14 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 3840E17ABE; Thu, 15 Jun 2000 15:35:13 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A5A61415DA for ; Thu, 15 Jun 2000 15:35:12 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA25427 for proteins-list@hgmp.mrc.ac.uk; Thu, 15 Jun 2000 15:35:12 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f X-Newsgroups: bionet.molbio.proteins From: "Alan A. Dombkowski" X-Sender: domski@seawolf.gpcc.itd.umich.edu Subject: Re: Engineering a disulfide bond In-Reply-To: <8fa30p$nb6$1@news.doit.wisc.edu> Message-ID: References: <8fa30p$nb6$1@news.doit.wisc.edu> MIME-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII Lines: 24 Date: Thu, 15 Jun 2000 10:34:37 -0400 X-Trace: news.itd.umich.edu 961079684 141.211.2.153 (Thu, 15 Jun 2000 10:34:44 EDT) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Larry, I may be of some help. Email me. Your email address seems to be invalid. On 9 May 2000, Larry wrote: > My thesis work involves studying the protein-protein interactions > between two enzymes. One of the interaction domains is a short 69 > amino acid coiled-coil motif. We've cloned and expressed this region > in E. coli but want to engineer a disulfide bond to "close" off the > loop (and make the motif more rigid). I know that disulfide bond formation > is subject to many variables (correct phi, psi angles, etc.). Is > there a computer or web-based program that will help me predict where > I can insert two cysteines to create this disulfide bond? Thanks. > > > > From owner-proteins@hgmp.mrc.ac.uk Thu Jun 15 22:30:23 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8FEA217B07; Thu, 15 Jun 2000 22:30:19 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 56B3717AEA; Thu, 15 Jun 2000 22:30:16 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 3CF42415DA for ; Thu, 15 Jun 2000 22:30:15 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA09030 for proteins-list@hgmp.mrc.ac.uk; Thu, 15 Jun 2000 22:30:14 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: czhu00@my-deja.com X-Newsgroups: bionet.molbio.proteins Subject: Phosphorylation labeling in baculovirus cells Date: Thu, 15 Jun 2000 21:20:41 GMT Organization: Deja.com - Before you buy. Lines: 13 Message-ID: <8ibhan$fsv$1@nnrp1.deja.com> X-Article-Creation-Date: Thu Jun 15 21:20:41 2000 GMT X-Http-User-Agent: Mozilla/4.0 (compatible; MSIE 5.0; Windows 98; DigExt) X-Http-Proxy: 1.1 x61.deja.com:80 (Squid/1.1.22) for client 128.218.127.170 X-MyDeja-Info: XMYDJUIDczhu00 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Anybody has any experience in 32P labeling of proteins expressed in baculovirus infected insect cells? How much [32P]orthophosphate should I use and for how long should the cells be labeled? Thanks in advance for your help. Chang Zhu Microbiology UCSF http://geocities.yahoo.com/virtualab/ Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Fri Jun 16 08:23:32 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B522717A82; Fri, 16 Jun 2000 08:23:31 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 3CB0417A5E; Fri, 16 Jun 2000 08:23:30 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id B7A11415DA for ; Fri, 16 Jun 2000 08:23:29 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA16114 for proteins-list@hgmp.mrc.ac.uk; Fri, 16 Jun 2000 08:23:29 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: jht63378@netspace.net.au X-Newsgroups: bionet.molbio.proteins Subject: Find out real info about real people 28411 Date: 16 Jun 2000 08:23:27 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 296 Message-ID: <000011a011d5$00007b40$00006efb@212.211.171.146> X-Trace: niobium.hgmp.mrc.ac.uk 961140208 16111 193.62.192.80 (16 Jun 2000 07:23:28 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 58EA517A5E; Fri, 16 Jun 2000 08:23:26 +0100 (BST) X-Received: from mail.lyche.no (mail.lyche.no [195.18.192.70]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 1C2CA415DA; Fri, 16 Jun 2000 08:23:23 +0100 (BST) X-Received: from 195.146.99.116 ([38.27.42.125]) by mail.lyche.no (Lotus SMTP MTA v4.6.2 (693.3 8-11-1998)) with SMTP id 41256900.002D77F7; Fri, 16 Jun 2000 09:16:43 +0100 X-To: X-Priority: 3 X-MSMail-Priority: Normal X-Errors-To: tvr738@winning.com X-Reply-To: jht63378@netspace.net.au To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk We have all heard about the vast amounts of information that an investigator can obtain about an individual. 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Please be aware that any disruption to our remove link prevents those that want to be removed from our database from being removed. mailto:nadapromos358@usa.com?subject=pleaseremovecyberpromo --- From owner-proteins@hgmp.mrc.ac.uk Fri Jun 16 08:46:05 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8A8B517A84; Fri, 16 Jun 2000 08:46:04 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 3F5E017A82; Fri, 16 Jun 2000 08:46:03 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 9EE4B415DA for ; Fri, 16 Jun 2000 08:46:02 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA16465 for proteins-list@hgmp.mrc.ac.uk; Fri, 16 Jun 2000 08:46:02 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: j.berges@qub.AC.UK ("John A. Berges") X-Newsgroups: bionet.molbio.proteins Subject: (NONE) Date: 16 Jun 2000 08:46:01 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 4 Message-ID: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; CHARSET=US-ASCII X-Trace: niobium.hgmp.mrc.ac.uk 961141561 16463 193.62.192.80 (16 Jun 2000 07:46:01 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 6589817A82; Fri, 16 Jun 2000 08:46:00 +0100 (BST) X-Received: from mailhub2.qub.ac.uk (jeremiah.qub.ac.uk [143.117.14.29]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 65B02415DA for ; Fri, 16 Jun 2000 08:45:59 +0100 (BST) X-Received: from fujin.qub.ac.uk by mailhub2.qub.ac.uk with SMTP-QUB (XT-PP) with ESMTP; Fri, 16 Jun 2000 07:45:50 +0000 X-Received: from eagle.qub.ac.uk (e7206.bc.qub.ac.uk [143.117.47.206]) by fujin.qub.ac.uk (8.8.8+Sun/8.8.8) with SMTP id IAA23273 for ; Fri, 16 Jun 2000 08:45:44 +0100 (BST) X-To: proteins@hgmp.mrc.ac.uk X-Priority: NORMAL X-Mailer: Simeon for Win32 Version 4.1.5 Build (43) X-Authentication: IMSP To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk subscribe proteins --- From owner-proteins@hgmp.mrc.ac.uk Fri Jun 16 19:59:59 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3A00A17AFE; Fri, 16 Jun 2000 19:59:59 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 14A6017AF3; Fri, 16 Jun 2000 19:59:58 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 8C855415DC for ; Fri, 16 Jun 2000 19:59:57 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA24968 for proteins-list@hgmp.mrc.ac.uk; Fri, 16 Jun 2000 19:59:56 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: guthrie@Hub.TCH.Harvard.edu ("Paul D. Guthrie") X-Newsgroups: bionet.molbio.proteins Subject: prokaryotic expression systems?? Date: 16 Jun 2000 19:59:55 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 23 Message-ID: <394A87C9.95469415@hub.tch.harvard.edu> Mime-Version: 1.0 Content-Type: text/plain; x-mac-creator=4D4F5353; x-mac-type=54455854; charset=us-ascii Content-Transfer-Encoding: 7BIT X-Trace: niobium.hgmp.mrc.ac.uk 961181996 24966 193.62.192.80 (16 Jun 2000 18:59:56 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id DB10017AF3; Fri, 16 Jun 2000 19:59:54 +0100 (BST) X-Received: from AREEL.TCH.HARVARD.EDU (areel.tch.harvard.edu [134.174.30.210]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 89B77415DC for ; Fri, 16 Jun 2000 19:59:50 +0100 (BST) X-Received: from hub.tch.harvard.edu ("port 1637"@* [134.174.42.240]) by Mail.TCH.Harvard.edu (PMDF V5.2-32 #32538) with ESMTP id <01JQOAY8GXXM92A0TP@Mail.TCH.Harvard.edu> for proteins@net.bio.net; Fri, 16 Jun 2000 14:58:55 -0400 (EDT) X-To: proteins@net.bio.net X-Mailer: Mozilla 4.7 (Macintosh; I; PPC) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk We're looking to express and purify a human protein of about 120 amino acids in E. coli. Does anyone have any feelings, one way or the other, about the following systems? Clontech pPROTet.E Promega PinPoint Xa Stratagene pCAL The Stratagene system looks pretty attractive, but we'd like to hear from someone who's actually used one or all of these. Thanks for any advice!! Paul D. Guthrie Urology Research Lab Children's Hospital, Boston --- From owner-proteins@hgmp.mrc.ac.uk Mon Jun 19 18:38:38 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5AEA317AD2; Mon, 19 Jun 2000 18:38:37 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 8EDBE17AAC; Mon, 19 Jun 2000 18:38:35 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 8FBEE415DC for ; Mon, 19 Jun 2000 18:38:34 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id SAA10429 for proteins-list@hgmp.mrc.ac.uk; Mon, 19 Jun 2000 18:38:33 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "P. Moreno" X-Newsgroups: bionet.molbio.proteins Subject: Re: prokaryotic expression systems?? Date: Mon, 19 Jun 2000 19:29:59 +0200 Organization: Universitat de Valencia Lines: 49 Message-ID: <394E5896.F7275683@servidor.ibv.csic.es> References: <394A87C9.95469415@hub.tch.harvard.edu> Reply-To: pmoreno@ibv.csic.es Mime-Version: 1.0 Content-Type: multipart/mixed; boundary="------------A1CE40151A53CA16A778E3DF" X-Trace: peque.uv.es 961435857 13306 161.111.218.148 (19 Jun 2000 17:30:57 GMT) X-Complaints-To: postmaster@uv.es X-Mailer: Mozilla 4.7 [en] (Win98; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk This is a multi-part message in MIME format. --------------A1CE40151A53CA16A778E3DF Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit "Paul D. Guthrie" wrote: > We're looking to express and purify a human protein of about 120 amino > acids in E. coli. Does anyone have any feelings, one way or the other, > about the following systems? > > Clontech pPROTet.E > > Promega PinPoint Xa > > Stratagene pCAL > > The Stratagene system looks pretty attractive, but we'd like to hear > from someone who's actually used one or all of these. Thanks for any > advice!! > > Paul D. Guthrie > Urology Research Lab > Children's Hospital, Boston > > --- I have made it with pET system ( novagen ); and the result was well. --------------A1CE40151A53CA16A778E3DF Content-Type: text/x-vcard; charset=us-ascii; name="pmoreno.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for P. Moreno Content-Disposition: attachment; filename="pmoreno.vcf" begin:vcard n:; x-mozilla-html:FALSE adr:;;;;;; version:2.1 end:vcard --------------A1CE40151A53CA16A778E3DF-- From owner-proteins@hgmp.mrc.ac.uk Tue Jun 20 14:18:10 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 6443017B15; Tue, 20 Jun 2000 14:18:01 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7AB1617AA2; Tue, 20 Jun 2000 14:17:57 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id E972C415DD; Tue, 20 Jun 2000 14:17:56 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA02878; Tue, 20 Jun 2000 14:17:56 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Ian Harper X-Newsgroups: bionet.cellbiol,bionet.cellbiol.cytonet,bionet.molbio.proteins,bionet.molbio.methds-reagnts Subject: Help with Taxol preps Date: Tue, 20 Jun 2000 23:13:48 +1000 Organization: Monash Uni Lines: 27 Distribution: world Message-ID: <394F6E0C.C9F8791E@sci.monash.edu.au> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: towncrier.cc.monash.edu.au 961507074 12371 130.194.10.139 (20 Jun 2000 13:17:54 GMT) X-Complaints-To: abuse@monash.edu.au X-Mailer: Mozilla 4.7 [en] (WinNT; I) X-Accept-Language: en To: cellbiol@hgmp.mrc.ac.uk, cytonet@hgmp.mrc.ac.uk, methods@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi, A student of ours is using Taxol to disrupt the cytoskeleton, and has started getting unexpected preciptations forming in the medium after switching from a pharmaceutical grade to the "sigma" powder. A stock is made up either in DMSO or Cremophor El (oil based I think) + Ethanol ( as suggested by supplier), but when the stock is diluted 0.25% vol to the aqueous, we see needle-shaped crystal forming as taxol goes out of solution. Ideas, and protocols would be greatly appreciated, plz email to me: Ian.Harper@sci.monash.edu.au Thanks Ian. ><1><1><1><1><1><1><1>< Ian Harper Confocal Imaging Facility Dept Biological Sciences Monash University Wellington Rd, Clayton VIC, Australia Tel: +61-3-99055635, Fax: +61-3-99055613 Email: Ian.Harper@sci.monash.edu.au ><1><1><1><1><1><1><1>< From owner-proteins@hgmp.mrc.ac.uk Tue Jun 20 17:00:30 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 8B69117AC4; Tue, 20 Jun 2000 17:00:29 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 87C9417A8A; Tue, 20 Jun 2000 17:00:26 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 41245415DA for ; Tue, 20 Jun 2000 17:00:25 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id RAA00458 for proteins-list@hgmp.mrc.ac.uk; Tue, 20 Jun 2000 17:00:24 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Tina " X-Newsgroups: bionet.molbio.proteins Subject: HELP Lines: 8 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2919.6600 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2919.6600 Message-ID: Date: Tue, 20 Jun 2000 15:00:00 GMT X-Complaints-To: abuse@news.chello.at X-Trace: news.chello.at 961513200 212.17.107.159 (Tue, 20 Jun 2000 17:00:00 MET DST) Organization: Chello Austria To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Does a transport mechanism exist in the mitochondrial membran, that is spezific for citrate, succinate and malate ? Where can I find some information? thx for your help, sigrid From owner-proteins@hgmp.mrc.ac.uk Wed Jun 21 08:37:37 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 7927717ACD; Wed, 21 Jun 2000 08:37:36 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id C3A2717AC9; Wed, 21 Jun 2000 08:37:34 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 29EFD415DA for ; Wed, 21 Jun 2000 08:37:34 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA19855 for proteins-list@hgmp.mrc.ac.uk; Wed, 21 Jun 2000 08:37:33 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: cruiseafree9@indiatimes.com X-Newsgroups: bionet.molbio.proteins Subject: FREE: Tropical Caribbean Trip & Deluxe Hotel Date: 21 Jun 2000 08:37:32 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 24 Message-ID: <20000621073729.4922F415DA@mercury.hgmp.mrc.ac.uk> X-Trace: niobium.hgmp.mrc.ac.uk 961573053 19853 193.62.192.80 (21 Jun 2000 07:37:33 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7581A17AC9; Wed, 21 Jun 2000 08:37:30 +0100 (BST) X-Received: from inetsrv0.sohard.ch (dnssohard1.sohard.ch [194.230.67.5]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 4922F415DA for ; Wed, 21 Jun 2000 08:37:29 +0100 (BST) X-Received: from iduhah9 (98-pool1.ras10.midea.agisdial.net [209.14.80.98]) by inetsrv0.sohard.ch with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2650.21) id NK9KKKBF; Wed, 21 Jun 2000 09:35:43 +0200 X-To: vacationnow9@hotmail.com To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk


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--- From owner-proteins@hgmp.mrc.ac.uk Wed Jun 21 10:45:39 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id BFB9117B34; Wed, 21 Jun 2000 10:45:38 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 079EB17B33; Wed, 21 Jun 2000 10:45:36 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 519D6415DA for ; Wed, 21 Jun 2000 10:45:36 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id KAA21462 for proteins-list@hgmp.mrc.ac.uk; Wed, 21 Jun 2000 10:45:35 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Andreas Jerlich X-Newsgroups: bionet.molbio.proteins Subject: Tryptophans in Hemoglobin ?? Date: Wed, 21 Jun 2000 11:29:54 +0200 Organization: Karl-Franzens-Universitaet Graz, Austria Lines: 13 Message-ID: <39508B12.861714E5@gmx.at> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: jerry.kfunigraz.ac.at 961579955 216 143.50.24.106 (21 Jun 2000 09:32:35 GMT) X-Complaints-To: usenet@news.kfunigraz.ac.at X-Mailer: Mozilla 4.51 [en] (Win95; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I am looking for the number of Tryptophans in Hemoglobin. Any information appreciated (URL, publication ...) Thanks in advance, Andreas -- ******************************************************* Andreas Jerlich - IMBM - Uni Graz - Austria A.J.Honest@gmx.at http://www-ang.kfunigraz.ac.at/~jerlich ******************************************************* From owner-proteins@hgmp.mrc.ac.uk Wed Jun 21 13:20:19 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 3FC4E17AF0; Wed, 21 Jun 2000 13:20:19 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id AC01917AE4; Wed, 21 Jun 2000 13:20:17 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 18205415DC for ; Wed, 21 Jun 2000 13:20:17 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id NAA23335 for proteins-list@hgmp.mrc.ac.uk; Wed, 21 Jun 2000 13:20:16 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Kresten X-Newsgroups: bionet.molbio.proteins Subject: Re: redox buffer novice Date: Wed, 21 Jun 2000 12:10:09 GMT Organization: Deja.com - Before you buy. Lines: 47 Message-ID: <8iqbap$sa$1@nnrp1.deja.com> References: <39480278.83B239E8@rocketmail.com> X-Article-Creation-Date: Wed Jun 21 12:10:09 2000 GMT X-Http-User-Agent: Mozilla/4.0 (compatible; MSIE 5.0; Windows 98; DigExt) X-Http-Proxy: 1.1 x61.deja.com:80 (Squid/1.1.22) for client 130.226.182.218 X-MyDeja-Info: XMYDJUIDkresten To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk > > I am studying the redox properties of my enzyme using various redox > buffers (GSH/GSSG; cysteine:cystine; DTTred/DTToxid). Most of the > protocols that have described in the literature for these buffers > don't > describe whether these buffers were deoxygenated prior to use; or > whether any precautions were taken to prevent oxidation during the > assay. Are there any protocols for redox buffers that take the effect > of oxygen into account ? I usually add a bit of EDTA (preventing metal-ion catalyzed oxidation) and purge extensively with Argon. > > One more question: the active site cysteines of may enzyme may become > oxidised to sulfinic or sulfenic acids ... how could I test this ? Ellman? Sequencing? IEF? Pyridine-ethylation and Amino acid analysis? ... HTH Kresten > > Thank you for your time, > > Che Pillay > Dept. of Biochemsitry > University of Natal > South Africa > pillayche@nu.ac.za > > --- > -- The address kresten@my-dejanews.com is for spambots only. Please mail me at LysLeuLeu@crc.dk transforming the pre@-part into my initials. Kresten Lindorff Larsen, Dept. Yeast Genetics Sent via Deja.com http://www.deja.com/ Before you buy. From owner-proteins@hgmp.mrc.ac.uk Wed Jun 21 14:31:34 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 72C0B17B2F; Wed, 21 Jun 2000 14:31:33 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id AF90D17B27; Wed, 21 Jun 2000 14:31:31 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 1EECA415DE for ; Wed, 21 Jun 2000 14:31:31 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA24161 for proteins-list@hgmp.mrc.ac.uk; Wed, 21 Jun 2000 14:31:30 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Matt Hicks" X-Newsgroups: bionet.molbio.proteins Subject: Re: Tryptophans in Hemoglobin ?? Date: Wed, 21 Jun 2000 14:24:16 +0100 Organization: University of Sussex Lines: 20 Message-ID: <8iqfoc$t8s$1@ames.central.susx.ac.uk> References: <39508B12.861714E5@gmx.at> X-Trace: ames.central.susx.ac.uk 961593932 29980 139.184.161.203 (21 Jun 2000 13:25:32 GMT) X-Complaints-To: usenet@newshost.central.susx.ac.uk X-Newsreader: Microsoft Outlook Express 4.72.3110.1 X-MimeOLE: Produced By Microsoft MimeOLE V4.72.3110.3 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk look at http://srs6.ebi.ac.uk/ to get sequences good luck Andreas Jerlich wrote in message <39508B12.861714E5@gmx.at>... >I am looking for the number of Tryptophans in Hemoglobin. Any >information appreciated (URL, publication ...) > >Thanks in advance, Andreas > >-- >******************************************************* > Andreas Jerlich - IMBM - Uni Graz - Austria > A.J.Honest@gmx.at > http://www-ang.kfunigraz.ac.at/~jerlich >******************************************************* > > From owner-proteins@hgmp.mrc.ac.uk Wed Jun 21 15:14:52 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id AEA8417AF0; Wed, 21 Jun 2000 15:14:51 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id EA99817A9B; Wed, 21 Jun 2000 15:14:49 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id EC5DA415DC for ; Wed, 21 Jun 2000 15:14:48 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA24817 for proteins-list@hgmp.mrc.ac.uk; Wed, 21 Jun 2000 15:14:48 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Chris LaRosa X-Newsgroups: bionet.molbio.proteins Subject: Re: Tryptophans in Hemoglobin ?? Date: Wed, 21 Jun 2000 09:12:40 -0500 Organization: University of Nebraska-Lincoln Lines: 11 Message-ID: <3950CD57.5C3D73FE@biocomp.unl.edu> References: <39508B12.861714E5@gmx.at> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.72 [en] (Win95; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Andreas Jerlich wrote: > > I am looking for the number of Tryptophans in Hemoglobin. Any > information appreciated (URL, publication ...) > > have you tried the GenBank/Embl/PIR Swiss-prot data bases??? You can get the sequences of any hemoglogin and using a huge variety of programs...count the number of each amino acid. From owner-proteins@hgmp.mrc.ac.uk Thu Jun 22 02:20:58 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 1BFF517A9C; Thu, 22 Jun 2000 02:20:57 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id A204917A5B; Thu, 22 Jun 2000 02:20:56 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id BB3F0415DC for ; Thu, 22 Jun 2000 02:20:55 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id CAA11388 for proteins-list@hgmp.mrc.ac.uk; Thu, 22 Jun 2000 02:20:55 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Scott Craig X-Newsgroups: bionet.molbio.proteins Subject: Protein extraction buffer Date: Thu, 22 Jun 2000 11:10:28 +1000 Organization: School of Biotechnology, University of New South Wales Lines: 14 Message-ID: <39516784.21F58494@student.unsw.edu.au> Mime-Version: 1.0 Content-Type: text/html; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: mirv.comms.unsw.edu.au 961636272 26505 149.171.144.202 (22 Jun 2000 01:11:12 GMT) X-Complaints-To: abuse@unsw.edu.au X-Mailer: Mozilla 4.72 [en] (Win95; U) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hi All,
I am after a recipe or reference for a protein extraction buffer similar to Pierce's Bper. I have been using Bper with good results but find it a bit expensive for routine use.
Any alternatives/ideas would be greatly appreciated.

--
Scott Craig
Dept. of Biotechnology
University of New South Wales
Sydney, Australia
  From owner-proteins@hgmp.mrc.ac.uk Fri Jun 23 15:16:17 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 0C4E517ACC; Fri, 23 Jun 2000 15:16:16 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id AAD6D17A68; Fri, 23 Jun 2000 15:16:14 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 5A9F1415CF for ; Fri, 23 Jun 2000 15:16:13 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA16932 for proteins-list@hgmp.mrc.ac.uk; Fri, 23 Jun 2000 15:16:12 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Bassie X-Newsgroups: bionet.molbio.proteins Subject: Dissolving gelatin Date: Fri, 23 Jun 2000 16:16:24 +0200 Organization: Vereniging Kabelfoongebruikersgroep Lines: 11 Message-ID: <39537137.2EABB1D5@kabelfoon.nl> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: news.kabelfoon.nl 961769766 49306 212.153.112.166 (23 Jun 2000 14:16:06 GMT) X-Complaints-To: abuse@kabelfoon.nl X-Mailer: Mozilla 4.08 [en] (Win95; I) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Hello all, I was wondering if the gelation of a protein (like gelatin) can be reversed. I did some experiments and found that after drying it or freeze/ melt you still get small particles instead of dissolved protein. I know that using a protease you can dissolve it but then the protein is distroyed completely, I would like to have intact redissolved gelatin. ---=== Bassie ===--- From owner-proteins@hgmp.mrc.ac.uk Fri Jun 23 15:59:01 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5834017B0A; Fri, 23 Jun 2000 15:59:00 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 3687B17AA2; Fri, 23 Jun 2000 15:58:58 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id A69EA415D4 for ; Fri, 23 Jun 2000 15:58:56 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA17466 for proteins-list@hgmp.mrc.ac.uk; Fri, 23 Jun 2000 15:58:56 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: raman@bragg.bio.uci.edu (CS Raman) X-Newsgroups: bionet.molbio.proteins Subject: POSTDOCTORAL POSITIONS IN STRUCTURAL BIOLOGY Date: 23 Jun 2000 15:58:55 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 37 Message-ID: <200006231457.OAA131668@hemebase.bio.uci.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 961772335 17464 193.62.192.80 (23 Jun 2000 14:58:55 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 727A717B24; Fri, 23 Jun 2000 15:58:46 +0100 (BST) X-Received: from bragg.bio.uci.edu (bragg.bio.uci.edu [128.195.151.87]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D29C7415D4 for ; Fri, 23 Jun 2000 15:58:44 +0100 (BST) X-Received: from hemebase.bio.uci.edu (hemebase.bio.uci.edu [128.195.151.52]) by bragg.bio.uci.edu (8.9.3/) with ESMTP id HAA31089 for ; Fri, 23 Jun 2000 07:58:43 -0700 (PDT) X-Received: (from raman@localhost) by hemebase.bio.uci.edu (8.9.3/) id OAA131668 for proteins@net.bio.net; Fri, 23 Jun 2000 14:57:38 GMT X-To: proteins@net.bio.net X-Mailer: ELM [version 2.5 PL3] To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk POSTDOCTORAL POSITIONS IN STRUCTURAL BIOLOGY Two positions are available in the newly established Structural Biology Center at the University of Texas. Highly motivated individuals are sought to pursue research in an active and collaborative environment. One position will focus on x-ray crystallographic studies of the proteins involved in the nitric oxide signaling pathway (Cell 95: 939-950 [1998]). Another position is geared towards solving structures of novel proteins in transcriptional silencing. Diffraction-quality crystals and/or optimized protein expression systems are in hand to provide a head-start on these projects. Both projects also present the opportunity for pursuing biochemical and functional studies in parallel with the structural analysis. The laboratory is equipped with state-of-the-art facilites including a Rigaku generator, RAXIS IV++ detectors, Osmic mirrors, cryo-cooling, and equipment for all aspects of protein purification, expression, and characterization. The applicant should have a Ph.D. in Biochemistry or Molecular Biophysics. Preference will be given to those candidates with training in macromolecular crystallography. However, those with research expertise in protein chemistry, molecular biology, or computational chemistry/biology are encouraged to apply. Interested candidates should send their CV along with e-mail addresses of three references to craman@uci.edu OR Dr. C. S. Raman, Structural Biology Center, Dept. of Biochemistry and Molecular Biology, University of Texas - Houston Medical School, Suite 6.200, 6431 Fannin St, Houston, TX 77030. URL:deanweb.med.uth.tmc.edu. The University of Texas - Medical School in Houston is an Equal Employment/ Opportunity Employer. --- From owner-proteins@hgmp.mrc.ac.uk Sun Jun 25 03:57:11 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id C418917B1B; Sun, 25 Jun 2000 03:57:10 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 9838217A9C; Sun, 25 Jun 2000 03:57:09 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 1C4F0415D4 for ; Sun, 25 Jun 2000 03:57:09 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id DAA29955 for proteins-list@hgmp.mrc.ac.uk; Sun, 25 Jun 2000 03:57:08 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: sudha@scripps.edu (Sudha Veeraraghavan) X-Newsgroups: bionet.molbio.proteins Subject: Postdoctoral Positions: Protein NMR Date: 25 Jun 2000 03:57:07 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 52 Message-ID: Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Trace: niobium.hgmp.mrc.ac.uk 961901828 29953 193.62.192.80 (25 Jun 2000 02:57:08 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 8B86517A9C; Sun, 25 Jun 2000 03:57:06 +0100 (BST) X-Received: from relay1.scripps.edu (relay1.scripps.edu [137.131.200.29]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 286F0415D4 for ; Sun, 25 Jun 2000 03:57:05 +0100 (BST) X-Received: from bianca.scripps.edu (bianca.scripps.edu [137.131.252.86]) by relay1.scripps.edu (8.9.3/TSRI-3.2.0r) with ESMTP id TAA17364 for ; Sat, 24 Jun 2000 19:57:03 -0700 (PDT) X-Received: from localhost (sudha@localhost) by bianca.scripps.edu (8.9.2/TSRI-3.0.1) with SMTP id TAA16041 for ; Sat, 24 Jun 2000 19:57:02 -0700 (PDT) X-Sender: sudha@bianca X-To: proteins@net.bio.net To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Postdoctoral Positions Structural Biology Center University of Texas-Houston Medical School Two postdoctoral positions are available immediately to solve protein structures using modern NMR methods. Research focus is to elucidate the molecular basis for cell cycle regulation. One project involves studies on a novel protein that regulates oocyte maturation; another involves investigating a kinase that plays a key role in cell proliferation. Fellows will study the solution structures and dynamics of proteins or protein fragments using hetero- and homonuclear multidimensional gradient NMR techniques on a 600 MHz Avance NMR spectrometer. The projects will also incorporate proteomics approaches to identify and characterize interacting proteins. Candidates who are competent in macromolecular NMR methods and solving structures or with a strong background in pulse program development and interest in learning to solve structures are preferred. Interested candidates should e-mail their CV with the names, addresses, and e-mail of three references to sudha@scripps.edu or mail to Dr. Sudha Veeraraghavan Structural Biology Center Department of Biochemistry and Molecular Biology University of Texas-Houston Medical School 6431 Fannin Street Houston, TX 77030 ***The UT-Houston Medical School is an Equal Opportunity/Affirmative Action Employer *** !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! ************************************************************************* Sudha Veeraraghavan, Ph.D. E-Mail: sudha@scripps.edu Department of Molecular Biology TPC15 The Scripps Research Institute Office: (858) 784-8824 10550 N. Torrey Pines Rd. Harper Lab: (858) 784-9858 La Jolla, CA 92037 FAX : (858) 784-8895 ^^^^ ************************************************************************** !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! --- From owner-proteins@hgmp.mrc.ac.uk Mon Jun 26 08:45:28 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 9A03317ACA; Mon, 26 Jun 2000 08:45:27 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 5BBFD17A9F; Mon, 26 Jun 2000 08:45:26 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D3FCE415CF for ; Mon, 26 Jun 2000 08:45:25 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA29003 for proteins-list@hgmp.mrc.ac.uk; Mon, 26 Jun 2000 08:45:25 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Andreas Jerlich X-Newsgroups: bionet.molbio.proteins Subject: Re: Tryptophans in Hemoglobin ?? Date: Mon, 26 Jun 2000 09:34:13 +0200 Organization: Karl-Franzens-Universitaet Graz, Austria Lines: 22 Message-ID: <39570775.D8BFF140@gmx.at> References: <39508B12.861714E5@gmx.at> <3950CD57.5C3D73FE@biocomp.unl.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: jerry.kfunigraz.ac.at 962005015 24289 143.50.24.106 (26 Jun 2000 07:36:55 GMT) X-Complaints-To: usenet@news.kfunigraz.ac.at X-Mailer: Mozilla 4.51 [en] (Win95; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Thanks !!! Chris LaRosa wrote: > Andreas Jerlich wrote: > > > > I am looking for the number of Tryptophans in Hemoglobin. Any > > information appreciated (URL, publication ...) > > > > > have you tried the GenBank/Embl/PIR Swiss-prot data bases??? > You can get the sequences of any hemoglogin and using a huge variety of > programs...count the number of each amino acid. -- ******************************************************* Andreas Jerlich - IMBM - Uni Graz - Austria A.J.Honest@gmx.at http://www-ang.kfunigraz.ac.at/~jerlich ******************************************************* From owner-proteins@hgmp.mrc.ac.uk Mon Jun 26 15:06:29 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 43CD817B0E; Mon, 26 Jun 2000 15:06:28 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 9DC3B17ABD; Mon, 26 Jun 2000 15:06:26 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 24944415D5 for ; Mon, 26 Jun 2000 15:06:26 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id PAA04586 for proteins-list@hgmp.mrc.ac.uk; Mon, 26 Jun 2000 15:06:25 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: Chris LaRosa X-Newsgroups: bionet.molbio.proteins Subject: transmembrane protein prediction software Date: Mon, 26 Jun 2000 09:04:19 -0500 Organization: University of Nebraska-Lincoln Lines: 10 Message-ID: <395762E3.A39DF858@biocomp.unl.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.72 [en] (Win95; I) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk There are a number of transmembrane prediction routines on the web now, that supposedly are better than Kyte and Doolittle hydrophobicity plots. For instance, at www.expasy.ch there are about 6 routines. When I run a protein on them I get 2 that suggest that there is a transmembrane spanning region (DAS, tmpred2), the others do not. Not being a protein biochemist primarily, I wonder if some kind experts could comment on the utility of these routines and perhaps point me to some reviews of these new-fangled prediction routines? P. Christopher Larosa, PhD From owner-proteins@hgmp.mrc.ac.uk Mon Jun 26 22:24:40 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 77DE117B36; Mon, 26 Jun 2000 22:24:39 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id A234517B2D; Mon, 26 Jun 2000 22:24:37 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id DED7D415CF for ; Mon, 26 Jun 2000 22:24:36 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id WAA18512 for proteins-list@hgmp.mrc.ac.uk; Mon, 26 Jun 2000 22:24:35 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "None" X-Newsgroups: bionet.molbio.proteins Subject: Free Protocols` Date: Mon, 26 Jun 2000 14:18:47 -0700 Organization: MindSpring Enterprises Lines: 8 Message-ID: <8j8hnd$sqe$1@slb7.atl.mindspring.net> X-Server-Date: 26 Jun 2000 21:25:01 GMT X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I was surfing the Net today and found a site that has tons of protocols. The site is free and the protocols are all edited. They also list their editorial board and contributors - very impressive names! The protocols I have looked at are excellent; they are all in the same format and easily understood. Check it out if you need a protocol: www.bioprotocol.com Mitch V. From owner-proteins@hgmp.mrc.ac.uk Tue Jun 27 04:23:36 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 819EF17A60; Tue, 27 Jun 2000 04:23:33 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id C9E3617A60; Tue, 27 Jun 2000 04:23:29 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 1D113415D5; Tue, 27 Jun 2000 04:23:29 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id EAA22779; Tue, 27 Jun 2000 04:23:28 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Centriole" X-Newsgroups: bionet.cellbiol,bionet.cellbiol.cytonet,bionet.molbio.proteins,bionet.molbio.methds-reagnts References: <394F6E0C.C9F8791E@sci.monash.edu.au> Subject: Re: Help with Taxol preps Lines: 39 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2919.6700 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2919.6700 Message-ID: Date: Tue, 27 Jun 2000 03:23:23 GMT X-Complaints-To: abuse@home.net X-Trace: news1.rdc1.il.home.com 962076203 24.10.95.177 (Mon, 26 Jun 2000 20:23:23 PDT) Organization: @Home Network To: cellbiol@hgmp.mrc.ac.uk, cytonet@hgmp.mrc.ac.uk, methods@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I got the same thing happening with Nocodazole. At first I thought my cells were contaminated. The interesting thing is, it only seems to precipitate after more than 24 hours incubation. Before that, I can see no ppt microscopically. I wonder if the pH is an issue here because of that observation. In any event, the expected results still occur (i.e.. metaphase arrest with condensed chromosomes). "Ian Harper" wrote in message news:394F6E0C.C9F8791E@sci.monash.edu.au... > Hi, > A student of ours is using Taxol to disrupt the cytoskeleton, and has > started getting unexpected preciptations forming in the medium after > switching from a pharmaceutical grade to the "sigma" powder. A stock is > made up either in DMSO or Cremophor El (oil based I think) + Ethanol ( > as suggested by supplier), but when the stock is diluted 0.25% vol to > the aqueous, we see needle-shaped crystal forming as taxol goes out of > solution. > Ideas, and protocols would be greatly appreciated, > plz email to me: Ian.Harper@sci.monash.edu.au > > Thanks > > Ian. > > ><1><1><1><1><1><1><1>< > Ian Harper > Confocal Imaging Facility > Dept Biological Sciences > Monash University > Wellington Rd, Clayton > VIC, Australia > Tel: +61-3-99055635, Fax: +61-3-99055613 > Email: Ian.Harper@sci.monash.edu.au > ><1><1><1><1><1><1><1>< > > From owner-proteins@hgmp.mrc.ac.uk Tue Jun 27 05:14:03 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id B78FC17B34; Tue, 27 Jun 2000 05:14:02 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 8142717B09; Tue, 27 Jun 2000 05:14:01 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id AB958415D5 for ; Tue, 27 Jun 2000 05:14:00 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id FAA23385 for proteins-list@hgmp.mrc.ac.uk; Tue, 27 Jun 2000 05:14:00 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Ben" X-Newsgroups: bionet.molbio.proteins Subject: kinase Lines: 6 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 Message-ID: Date: Tue, 27 Jun 2000 04:13:58 GMT X-Trace: carnaval.risq.qc.ca 962079238 206.167.180.162 (Tue, 27 Jun 2000 00:13:58 EDT) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I want to know the kinase involved in phosphorylation of the enzyme studied (cytochrome P450c17), is there a technique by which I could "trap" this specific kinase ?? From owner-proteins@hgmp.mrc.ac.uk Tue Jun 27 06:00:56 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 618F317B0B; Tue, 27 Jun 2000 06:00:55 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id CF69017B06; Tue, 27 Jun 2000 06:00:53 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 127B2415D4 for ; Tue, 27 Jun 2000 06:00:52 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id GAA23965 for proteins-list@hgmp.mrc.ac.uk; Tue, 27 Jun 2000 06:00:52 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Neil Saunders" X-Newsgroups: bionet.molbio.proteins Subject: ArchaeaWeb-announce Date: Tue, 27 Jun 2000 14:57:51 +1000 Organization: UNSW Lines: 26 Message-ID: <8j9c8d$4ld$1@mirv.comms.unsw.edu.au> Reply-To: "Neil Saunders" X-Trace: mirv.comms.unsw.edu.au 962081869 4781 149.171.171.96 (27 Jun 2000 04:57:49 GMT) X-Complaints-To: abuse@unsw.edu.au X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2919.6600 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2919.6600 To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk http://www.crosswinds.net/~nfws/archaea/index.htm ArchaeaWeb is a new website aimed at collecting together web-related archaea and extremophile information in one place. Current features include a facility to search the PDB for archaeal protein structures, links to archaeal genome projects, links to archaea research labs, articles and the current weeks PubMed and GenBank submissions for archaeal papers and sequences. Comments welcome. Neil Saunders -- School of Microbiology & Immunology, University of New South Wales, Sydney 2052, Australia Ph: +61 2 9385 2093 Fx: +61 2 9385 1591 email: neil.saunders@unsw.edu.au WWW: http://www.crosswinds.net/~nfws/index.htm From owner-proteins@hgmp.mrc.ac.uk Tue Jun 27 14:59:15 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 97E2D17B3E; Tue, 27 Jun 2000 14:59:14 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 47B1517B34; Tue, 27 Jun 2000 14:59:13 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id AFAAA415DC for ; Tue, 27 Jun 2000 14:59:12 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id OAA00691 for proteins-list@hgmp.mrc.ac.uk; Tue, 27 Jun 2000 14:59:12 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: stgoif@dgdfg.com X-Newsgroups: maus.freizeit.f+sf,bionet.molbio.proteins Subject: C,A.B-L,E.B-O,X. -I`N,S.T-R,U.C-T`I,O.N-S, .B-U,I.L`D, .C-A,B.L-E` ,B.O-X,...`I.s, f-r,e.e- ,C.a-b`le, .P-os,s.i-b,l.e`?,.-,.-,.-.,.,,.......,....... 1643 Lines: 62 Message-ID: <1q265.30014$dP3.9752@news-reader.ntrnet.net> Date: Tue, 27 Jun 2000 13:58:21 GMT X-Complaints-To: abuse@ntrnet.net X-Trace: news-reader.ntrnet.net 962114301 63.24.129.164 (Tue, 27 Jun 2000 09:58:21 EDT) To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk L.EGAL C.A.B.L.E TV DE-S.C.R.A.M.B.L.E.R Want to watch Sporting Events?--Movies?--Pay-Per-View?? *This is the Famous R-O Shack TV D.e.s.c.r.a.m.b.l.e.r You can assemble it from R,a,d,i,o S,h,a,c,k parts for about $12 to $15. 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(Florida residents include 7% Florida State Sales Tax) (All orders outside the U.S.A. add $5.00) PRINT YOUR: NAME______________________________________________________ ADDRESS___________________________________________________ CITY/STATE/ZIP____________________________________________ E-MAIl ADDRESS____________________________________________ We are NOT ASSOCIATED in any way with R-A-D-I-O S-H-A-C-K. 4rwerwer24534ty5tgertrwetertertretrtre 3r345345345435435345435345435 whgoszlzqvgwivfbuzlxxvugvubodiljsplemiuxqqsxvsnmgfflxzrijnvhqsi From owner-proteins@hgmp.mrc.ac.uk Wed Jun 28 02:20:57 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 05A4E17AF0; Wed, 28 Jun 2000 02:20:52 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 44E5E17AAD; Wed, 28 Jun 2000 02:20:48 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 6C3CF415DC; Wed, 28 Jun 2000 02:20:47 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id CAA17389; Wed, 28 Jun 2000 02:20:46 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: stapletl@mail.nih.gov X-Newsgroups: bionet.cellbiol,bionet.cellbiol.cytonet,bionet.molbio.proteins,bionet.molbio.methds-reagnts Subject: Re: Help with Taxol preps Date: 27 Jun 2000 17:42:06 -0700 Organization: Extra Newsguy News Service [http://extra.newsguy.com] Lines: 46 Message-ID: <8jbhku$1cjb@edrn.newsguy.com> References: <394F6E0C.C9F8791E@sci.monash.edu.au> To: cellbiol@hgmp.mrc.ac.uk, cytonet@hgmp.mrc.ac.uk, methods@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk Have either of you considered checking if you are precipitating excess kinesin or other microtubule binding proteins (like on a Western)? Are you sure that you are not seeing inclusion bodies? In article , "Centriole" says... > >I got the same thing happening with Nocodazole. At first I thought my cells >were contaminated. The interesting thing is, it only seems to precipitate >after more than 24 hours incubation. Before that, I can see no ppt >microscopically. I wonder if the pH is an issue here because of that >observation. In any event, the expected results still occur (i.e.. >metaphase arrest with condensed chromosomes). > > >"Ian Harper" wrote in message >news:394F6E0C.C9F8791E@sci.monash.edu.au... >> Hi, >> A student of ours is using Taxol to disrupt the cytoskeleton, and has >> started getting unexpected preciptations forming in the medium after >> switching from a pharmaceutical grade to the "sigma" powder. A stock is >> made up either in DMSO or Cremophor El (oil based I think) + Ethanol ( >> as suggested by supplier), but when the stock is diluted 0.25% vol to >> the aqueous, we see needle-shaped crystal forming as taxol goes out of >> solution. >> Ideas, and protocols would be greatly appreciated, >> plz email to me: Ian.Harper@sci.monash.edu.au >> >> Thanks >> >> Ian. >> >> ><1><1><1><1><1><1><1>< >> Ian Harper >> Confocal Imaging Facility >> Dept Biological Sciences >> Monash University >> Wellington Rd, Clayton >> VIC, Australia >> Tel: +61-3-99055635, Fax: +61-3-99055613 >> Email: Ian.Harper@sci.monash.edu.au >> ><1><1><1><1><1><1><1>< >> >> > > From owner-proteins@hgmp.mrc.ac.uk Wed Jun 28 18:16:34 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 0E22D17AD1; Wed, 28 Jun 2000 18:16:29 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id E83CB17A82; Wed, 28 Jun 2000 18:16:25 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 66EAC415DA; Wed, 28 Jun 2000 18:16:25 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id SAA29239; Wed, 28 Jun 2000 18:16:24 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: "Centriole" X-Newsgroups: bionet.cellbiol,bionet.cellbiol.cytonet,bionet.molbio.proteins,bionet.molbio.methds-reagnts References: <394F6E0C.C9F8791E@sci.monash.edu.au> <8jbhku$1cjb@edrn.newsguy.com> Subject: Re: Help with Taxol preps Lines: 62 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2919.6700 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2919.6700 Message-ID: Date: Wed, 28 Jun 2000 17:16:22 GMT X-Complaints-To: abuse@home.net X-Trace: news1.rdc1.il.home.com 962212582 24.10.95.177 (Wed, 28 Jun 2000 10:16:22 PDT) Organization: @Home Network To: cellbiol@hgmp.mrc.ac.uk, cytonet@hgmp.mrc.ac.uk, methods@hgmp.mrc.ac.uk, proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I only see the ppt in the medium, not in the cells. The cells behave exactly as I expect them to in response to the nocodazole. After some time, there is a lot of the ppt. Much more than I would expect from cellular kinesin precipitation. wrote in message news:8jbhku$1cjb@edrn.newsguy.com... > Have either of you considered checking if you are precipitating excess kinesin > or other microtubule binding proteins (like on a Western)? Are you sure that > you are not seeing inclusion bodies? > > In article , "Centriole" says... > > > >I got the same thing happening with Nocodazole. At first I thought my cells > >were contaminated. The interesting thing is, it only seems to precipitate > >after more than 24 hours incubation. Before that, I can see no ppt > >microscopically. I wonder if the pH is an issue here because of that > >observation. In any event, the expected results still occur (i.e.. > >metaphase arrest with condensed chromosomes). > > > > > >"Ian Harper" wrote in message > >news:394F6E0C.C9F8791E@sci.monash.edu.au... > >> Hi, > >> A student of ours is using Taxol to disrupt the cytoskeleton, and has > >> started getting unexpected preciptations forming in the medium after > >> switching from a pharmaceutical grade to the "sigma" powder. A stock is > >> made up either in DMSO or Cremophor El (oil based I think) + Ethanol ( > >> as suggested by supplier), but when the stock is diluted 0.25% vol to > >> the aqueous, we see needle-shaped crystal forming as taxol goes out of > >> solution. > >> Ideas, and protocols would be greatly appreciated, > >> plz email to me: Ian.Harper@sci.monash.edu.au > >> > >> Thanks > >> > >> Ian. > >> > >> ><1><1><1><1><1><1><1>< > >> Ian Harper > >> Confocal Imaging Facility > >> Dept Biological Sciences > >> Monash University > >> Wellington Rd, Clayton > >> VIC, Australia > >> Tel: +61-3-99055635, Fax: +61-3-99055613 > >> Email: Ian.Harper@sci.monash.edu.au > >> ><1><1><1><1><1><1><1>< > >> > >> > > > > > From owner-proteins@hgmp.mrc.ac.uk Wed Jun 28 19:06:03 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id DF54817B17; Wed, 28 Jun 2000 19:06:02 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 5999917B29; Wed, 28 Jun 2000 19:05:59 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id D974E415E0 for ; Wed, 28 Jun 2000 19:05:58 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id TAA29840 for proteins-list@hgmp.mrc.ac.uk; Wed, 28 Jun 2000 19:05:58 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: deepti X-Newsgroups: bionet.molbio.proteins Subject: MALDI references Date: Wed, 28 Jun 2000 14:02:33 -0400 Organization: Yale University Lines: 8 Message-ID: <395A3DB6.F086EA7D@yale.edu> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.6 (Macintosh; I; PPC) X-Accept-Language: en To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk I'm trying to get specific references related to molecular weight determinations, using MALDI, of intact proteins - specifically the upper limit of detection. I know in theory it is greater than 300KDa, but anyone know what's actually been tested? Thanks, Deepti From owner-proteins@hgmp.mrc.ac.uk Wed Jun 28 20:14:26 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 5909417B32; Wed, 28 Jun 2000 20:14:26 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 415DA17AD1; Wed, 28 Jun 2000 20:14:25 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id BC50E415DC for ; Wed, 28 Jun 2000 20:14:24 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id UAA00727 for proteins-list@hgmp.mrc.ac.uk; Wed, 28 Jun 2000 20:14:24 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: ayourfreetv6@indiatimes.com X-Newsgroups: bionet.molbio.proteins Subject: Get a FREE Deluxe Satellite T.V. System ... Date: 28 Jun 2000 20:14:23 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 31 Message-ID: <20000628191416.7E5C5415DC@mercury.hgmp.mrc.ac.uk> X-Trace: niobium.hgmp.mrc.ac.uk 962219663 725 193.62.192.80 (28 Jun 2000 19:14:23 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 7866017AD1; Wed, 28 Jun 2000 20:14:22 +0100 (BST) X-Received: from bsserver.bsgate.it (unknown [212.35.202.67]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id 7E5C5415DC for ; Wed, 28 Jun 2000 20:14:16 +0100 (BST) X-Received: from detroit-ip-8-238.dynamic.ziplink.net [209.206.122.238] (HELO liahs12) by bsserver.bsgate.it (AltaVista Mail V2.0/2.0 BL23 listener) id 0000_0079_395a_4d98_8403; Wed, 28 Jun 2000 21:10:16 +0200 X-To: satsystems@hotmail.com To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk FREE SATELLITE T.V. SYSTEM

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Click Here to be removed. --- From owner-proteins@hgmp.mrc.ac.uk Thu Jun 29 08:34:41 2000 Return-Path: Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 110) id 89E5417B6A; Thu, 29 Jun 2000 08:34:40 +0100 (BST) Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id 33F4917B69; Thu, 29 Jun 2000 08:34:39 +0100 (BST) Received: from niobium.hgmp.mrc.ac.uk (niobium [193.62.192.41]) by mercury.hgmp.mrc.ac.uk (Postfix) with ESMTP id 85F66415DC for ; Thu, 29 Jun 2000 08:34:38 +0100 (BST) Received: (from news@localhost) by niobium.hgmp.mrc.ac.uk (8.9.3+Sun/8.8.8) id IAA18028 for proteins-list@hgmp.mrc.ac.uk; Thu, 29 Jun 2000 08:34:37 +0100 (BST) X-Authentication-Warning: niobium.hgmp.mrc.ac.uk: news set sender to using -f From: qqqwww@21cn.com (joe) X-Newsgroups: bionet.molbio.proteins Subject: protein diffusing in SDS-PAGE Date: 29 Jun 2000 08:34:36 +0100 Organization: BIOSCI/MRC Human Genome Mapping Project Resource Centre Lines: 4 Message-ID: <20000629073430.EB0EA415DC@mercury.hgmp.mrc.ac.uk> Mime-Version: 1.0 Content-Type: text/plain; charset="GB2312" Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 962264077 18026 193.62.192.80 (29 Jun 2000 07:34:37 GMT) X-Complaints-To: news@net.bio.net X-Received: by mercury.hgmp.mrc.ac.uk (Postfix, from userid 6024) id B615E17B69; Thu, 29 Jun 2000 08:34:35 +0100 (BST) X-Received: from 21cn.com (unknown [202.104.32.245]) by mercury.hgmp.mrc.ac.uk (Postfix) with SMTP id EB0EA415DC for ; Thu, 29 Jun 2000 08:34:30 +0100 (BST) X-Received: from qiao([202.103.166.252]) by 21cn.com(JetMail 2.5.3.0) with SMTP id jm2395b6b27; Thu, 29 Jun 2000 07:33:45 -0000 X-Reply-To: qqqwww@21cn.com X-To: proteins@net.bio.net X-Mailer: FoxMail 3.0 beta 2 [cn] To: proteins@hgmp.mrc.ac.uk Sender: owner-proteins@hgmp.mrc.ac.uk Precedence: bulk proteins I use 15% or 18% discontinuous SDS-PAGE gels,run at 150V until the dye front reaches the end of the gel (usually 50 minutes) and stained by G-250.I always find the band low than 14 MW is diffused,Anyone has experience in that and could give some suggestion?Thank you very much --- From owner-proteins@hgmp.mrc.ac.uk Thu Jun 29 08:57:19 2000 Return-Path: