From owner-proteins@net.bio.net Wed Dec 01 07:45:23 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news.maxwell.syr.edu!logbridge.uoregon.edu!uwm.edu!biosci!biosci!not-for-mail From: Chun-Song Yang Newsgroups: bionet.molbio.proteins, Subject: Function of CIB Protein Date: 30 Nov 1999 23:22:49 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 27 Sender: daemon@net.bio.net Distribution: world Message-ID: <822ic9$lsr@net.bio.net> NNTP-Posting-Host: net.bio.net Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:464 Hello, We had found a calcium and integrin binding protein (CIB) interacting with a mitochondria outer membrane receptor in our yeast two hybrid screening. We noticed that this protein might be involved in calcium ion signalling. But could anybody out there tell us whether or not this protein is involved in protein transport especially in or out mitochondria? Thanks for your attention. With best regards, Chun-Song ______________O_________oO_____________oO______o_______oO___________________ YEAST bionet newsgroup see: http://www.bio.net/hypermail/YEAST/ YEAST e-mail: messages sent to yeast@net.bio.net subscribe: e-mail biosci-server@net.bio.net with: subscribe yeast unsubscribe: e-mail biosci-server@net.bio.net with: unsubscribe yeast YEAST on the WWW: http://genome-www.stanford.edu/Saccharomyces/VL-yeast.html problems with the YEAST newsgroup? E-mail the moderator: francis@cmmt.ubc.ca From owner-proteins@net.bio.net Wed Dec 01 10:15:24 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news.algonet.se!algonet!newsfeed.tli.de!do.de.uu.net!news-koe1.dfn.de!news-han1.dfn.de!news-ham1.dfn.de!post.student.mu-luebeck.de!not-for-mail From: "Lars Komorowski" Newsgroups: bionet.molbio.proteins Subject: Re: His-tag intruder Date: Wed, 1 Dec 1999 10:54:17 +0100 Organization: Med. Universitaet zu Luebeck Lines: 44 Message-ID: <822qqe$bjt$1@post.student.mu-luebeck.de> References: <38398CA3.998DC9B0@umh.es> NNTP-Posting-Host: wahnsinn.biochem.mu-luebeck.de X-Trace: post.student.mu-luebeck.de 944041614 11901 141.83.167.171 (1 Dec 1999 09:46:54 GMT) X-Complaints-To: usenet@student.mu-luebeck.de NNTP-Posting-Date: 1 Dec 1999 09:46:54 GMT 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 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:465 In fact it is the isomerase SLYD. It binds Ni. I have had the same problem and we sequenced the N-terminus of the contaminant. Lars Jesús Sanz schrieb in im Newsbeitrag: 38398CA3.998DC9B0@umh.es... > Dear all, > > I'm sorry if this is and old one, but I have just come across this > problem. > When trying to purify my protein from E. coli as a His-tag fusion, I end > up co-purifiying > a very abundant 25 Kda protein (according to SDS-PAGE and mass > spectrometry) > that elutes at the same imidazole concentrations. > My protein is found in inclusion bodies as well as in the soluble > extract, but the > contaminant is also found in the soluble and insoluble fractions. > > I carried out an amino acid analysis and found plenty of histidines and > Asx + Glx. > The database shows that an E. coli peptidyl-prolyl isomerase of the > FKBP type has almost > the same percent composition, (and these proteins have been found to > bind to Ni2+ > columns very tightly) but its Mw is only around 21 Kda. > > FYI, I'm using the pLEX system (Invitrogen) in the GI724 E. coli strain. > > If this subject is already in the FAQ, please let me know where can I > access to it. > Thank you very much > ------------------------ > Prof. J. M. Sanz > Centro de Biologia Molecular y Celular > Universidad Miguel Hernandez > E-mail: jmsanz@umh.es > WWW:http://cbmc.umh.es/jmsanz/jesussanz.htm > ---------------- > > > From owner-proteins@net.bio.net Wed Dec 01 13:25:20 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news.maxwell.syr.edu!colt.net!news-lond.gip.net!news.gsl.net!gip.net!knews.uk0.vbc.net!vbcnet-gb!194.207.0.9.MISMATCH!news2.uk1.vbc.net!not-for-mail From: "SelectScience" Newsgroups: bionet.molbio.proteins Subject: leave your views online Date: Wed, 1 Dec 1999 12:56:53 -0000 Organization: VBCnet GB Ltd Lines: 7 Message-ID: <944055139.294763@news.uk1.vbc.net> NNTP-Posting-Host: news.uk1.vbc.net Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable X-Trace: news2.uk0.vbc.net 944054597 2358 194.207.2.4 (1 Dec 1999 13:23:17 GMT) X-Complaints-To: abuse@vbc.net NNTP-Posting-Date: 1 Dec 1999 13:23:17 GMT X-Newsreader: Microsoft Outlook Express 4.72.3110.5 X-MimeOLE: Produced By Microsoft MimeOLE V4.72.3110.3 Cache-Post-Path: news.uk1.vbc.net!unknown@194.207.229.174 X-Cache: nntpcache 2.3.3 (see http://www.nntpcache.org/) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:466 Any comments about manufacturers or their products? Come to http://www.selectscience.net =20 and leave your views on-line Editor SelectScience From owner-proteins@net.bio.net Wed Dec 01 18:31:48 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!warwick!server5.netnews.ja.net!easynet-tele!easynet.net!newspeer1.nac.net!news.maxwell.syr.edu!nntp2.deja.com!nnrp1.deja.com!not-for-mail From: Kresten Newsgroups: bionet.molbio.proteins Subject: Re: His-tag intruder Date: Wed, 01 Dec 1999 17:43:09 GMT Organization: Deja.com - Before you buy. Lines: 56 Message-ID: <823mnc$lns$1@nnrp1.deja.com> References: <38398CA3.998DC9B0@umh.es> NNTP-Posting-Host: 130.226.183.6 X-Article-Creation-Date: Wed Dec 01 17:43:09 1999 GMT X-Http-User-Agent: Mozilla/4.5 [en] (Win95; I) X-Http-Proxy: 1.0 www2.crc.dk:3128 (Squid/2.1.RELEASE), 1.0 x39.deja.com:80 (Squid/1.1.22) for client 130.226.182.217, 130.226.183.6 X-MyDeja-Info: XMYDJUIDkresten Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:467 As a comment; I've heard (somewhere) about people co-purifying chaperones on Ni2+ -columns. The problems ware apparently solved by adding ATP - probably releasing a protein-chaperone complex. Kresten In article <38398CA3.998DC9B0@umh.es>, =?iso-8859-1?Q?Jes=FAs?= Sanz wrote: > Dear all, > > I'm sorry if this is and old one, but I have just come across this > problem. > When trying to purify my protein from E. coli as a His-tag fusion, I end > up co-purifiying > a very abundant 25 Kda protein (according to SDS-PAGE and mass > spectrometry) > that elutes at the same imidazole concentrations. > My protein is found in inclusion bodies as well as in the soluble > extract, but the > contaminant is also found in the soluble and insoluble fractions. > > I carried out an amino acid analysis and found plenty of histidines and > Asx + Glx. > The database shows that an E. coli peptidyl-prolyl isomerase of the > FKBP type has almost > the same percent composition, (and these proteins have been found to > bind to Ni2+ > columns very tightly) but its Mw is only around 21 Kda. > > FYI, I'm using the pLEX system (Invitrogen) in the GI724 E. coli strain. > > If this subject is already in the FAQ, please let me know where can I > access to it. > Thank you very much > ------------------------ > Prof. J. M. Sanz > Centro de Biologia Molecular y Celular > Universidad Miguel Hernandez > E-mail: jmsanz@umh.es > WWW:http://cbmc.umh.es/jmsanz/jesussanz.htm > ---------------- > > -- 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@net.bio.net Thu Dec 02 02:55:52 1999 Path: hgmp.mrc.ac.uk!warwick!fu-berlin.de!logbridge.uoregon.edu!news.u.washington.edu!not-for-mail From: "Kun Qian" Newsgroups: bionet.molbio.proteins Subject: Immunoprecipitation question Date: Wed, 1 Dec 1999 18:42:12 -0800 Organization: University of Washington Lines: 13 Message-ID: <824mgc$154c$1@nntp4.u.washington.edu> NNTP-Posting-Host: cs102-15.fsmodem.washington.edu X-Trace: nntp4.u.washington.edu 944102732 38028 (None) 140.142.17.37 X-Complaints-To: help@cac.washington.edu X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2615.200 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:468 Hi, I am planning to do something related to immunopricipitation. I am asking that whether I should frezze the tissue samples by liquid nitrogen immediately after they are taken or I can safely leave the tissue in ordinary freezer (-80) without protein degradation? Thanks a lot! Kun Qian Department of Pediatrics University of Washington Seattle, WA 98195 From owner-proteins@net.bio.net Thu Dec 02 09:58:10 1999 Path: hgmp.mrc.ac.uk!warwick!fu-berlin.de!news.ruhrgebiet.individual.net!news-koe1.dfn.de!uni-muenster.de!not-for-mail From: Kai Schmengler Newsgroups: bionet.molbio.proteins Subject: unexpected protein expression Date: Thu, 02 Dec 1999 11:01:14 +0100 Organization: Westfaelische Wilhelms-Universitaet Muenster, Germany Lines: 26 Sender: "Kai Schmengler" Message-ID: <3846436A.F95C1D81@uni-muenster.de> NNTP-Posting-Host: porto.uni-muenster.de Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Trace: redenix.uni-muenster.de 944128687 29508 128.176.253.97 (2 Dec 1999 09:58:07 GMT) X-Complaints-To: abuse@uni-muenster.de NNTP-Posting-Date: 2 Dec 1999 09:58:07 GMT X-Mailer: Mozilla 4.61 [en] (Win98; I) X-Accept-Language: en Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:469 Dear all! During a ligation something occured which I can’t interpret yet. I used the cloning vector pPCRScript(Cam) containing a part of a new target gene flanked by the restriction sites BamHI (5’) and XmaIII(3’). The expression vector (pET20b) with the original gene was also cut with BamHI and XmaIII, respectively. Both, the new target gene and the rest vector were gel eluted. Then ligation was performed using T4-DNA-Ligase. After transformation several clones were checked on mini expression level resulting in only few proteins with the right size and many which were 15 kDa smaller. Sequencing information confirmed that the expression vector for the smaller protein contains a stop codon due to the loss of 28 nucleotides from the 5’-end of the insert. On the other hand, the vector expressing the right protein misses 45 nucleotides after the terminal XmaIII restriction site, suggesting that those nucleotides where cut from the original vector during the ligation (fortunately that doesn’t harm us). Did anybody experience something similar? I read something about lyase activity of ligases. May this be the reason? Thank you for your help. Kai From owner-proteins@net.bio.net Thu Dec 02 14:07:30 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!warwick!news.nott.ac.uk!news.nottingham.ac.uk!NewsWatcher!user From: kevin.bailey@nottingham.ac.uk () Newsgroups: bionet.molbio.proteins Subject: Endoprotease Lys-c Followup-To: bionet.molbio.proteins Date: 2 Dec 1999 13:58:10 GMT Organization: ACS, The University of Nottingham Lines: 6 Message-ID: NNTP-Posting-Host: wpbjwk1.biochem.nottingham.ac.uk X-Trace: oyez.ccc.nottingham.ac.uk 944143090 29170 128.243.183.238 (2 Dec 1999 13:58:10 GMT) X-Complaints-To: usenet@news.nottingham.ac.uk NNTP-Posting-Date: 2 Dec 1999 13:58:10 GMT Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:470 Anyone know of a cheaper source of sequencing grade Lys-c enzyme than that currently available from Boehringer (Roche)? I used to get it from Promega but they apparently have discontinued it. Boehringer charge about 117 pounds for 15 microgrammes (three times the price weight for weight of Promega's old price). Any other suppliers out there (preferably with a UK base) ? From owner-proteins@net.bio.net Thu Dec 02 21:43:15 1999 Path: hgmp.mrc.ac.uk!warwick!fu-berlin.de!howland.erols.net!torn!nott!uottawa!not-for-mail From: Neil McKenna Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Western stripping Date: Thu, 02 Dec 1999 16:36:01 -0500 Organization: University of Ottawa, Ottawa, Canada Lines: 8 Message-ID: <3846E641.A1DA76B4@ottawa.com> NNTP-Posting-Host: 232sub32.dynamic.uottawa.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Mailer: Mozilla 4.5 [en] (Win95; I) X-Accept-Language: en Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3316 bionet.molbio.proteins:471 I have had difficulty finding an efficient way to strip chemiluminescent/fluorescent-detected Western blots. I recall using a B-mercaptoethanol solution and/or an SDS solution, but I can't recall the concentrations or the steps. Is there anyone out there who has a protocol? Many thanks Neil From owner-proteins@net.bio.net Fri Dec 03 07:16:08 1999 Path: hgmp.mrc.ac.uk!warwick!fu-berlin.de!news-fra1.dfn.de!news.Uni-Mainz.DE!not-for-mail From: Thorsten Burmester Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Coomassie staining of proteins on PVDF - The answer? Followup-To: bionet.molbio.methds-reagnts Date: Thu, 02 Dec 1999 17:58:40 +0100 Organization: Johannes Gutenberg-Universitaet Mainz, Germany Lines: 22 Message-ID: <3846A540.E90ACBF3@erfurt.thur.de> References: <3833CB4F.1A549E22@erfurt.thur.de> NNTP-Posting-Host: poseidon.biologie.uni-mainz.de Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: bambi.zdv.Uni-Mainz.DE 944153819 15922 134.93.162.103 (2 Dec 1999 16:56:59 GMT) X-Complaints-To: usenet@mail.uni-mainz.de NNTP-Posting-Date: 2 Dec 1999 16:56:59 GMT To: Thorsten Burmester X-Mailer: Mozilla 4.5 [en] (Win95; I) X-Accept-Language: en,de Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3327 bionet.molbio.proteins:472 I asked the following question: > After SDS-PAGE and Western transfer to PVDF I stained the proteins with > 0.1% Coomassie Blue in H20 dd. Destaining with 50% methanol. To my > surprise the band of interest was _not_ stained but remained white on a > light blue background. Other proteins were stained well as expected. Any > explanation? FYI: According to the Proteomics people I talked to this phenomenon may be related to a low lysine (K) content of the protein (lack of positive charges?). I my case about 2 %. King & Jukes average protein 7.5 %. Thanks for all answers. Thorsten F'up -- Thorsten Burmester thorsten@erfurt.thur.de From owner-proteins@net.bio.net Fri Dec 03 07:52:55 1999 Path: hgmp.mrc.ac.uk!warwick!fu-berlin.de!news-fra1.dfn.de!aconews.univie.ac.at!news.univie.ac.at!not-for-mail From: "Martin Offterdinger" Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: Western stripping Date: Fri, 3 Dec 1999 08:53:03 +0100 Organization: Vienna University, Austria Lines: 16 Message-ID: <827ssj$6e5s$1@www.univie.ac.at> References: <3846E641.A1DA76B4@ottawa.com> Reply-To: "Martin Offterdinger" NNTP-Posting-Host: grunt.imed1.akh-wien.ac.at X-Trace: www.univie.ac.at 944207571 211132 149.148.97.3 (3 Dec 1999 07:52:51 GMT) X-Complaints-To: news-adm@news.univie.ac.at NNTP-Posting-Date: 3 Dec 1999 07:52:51 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 Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3328 bionet.molbio.proteins:473 Neil McKenna schrieb in im Newsbeitrag: 3846E641.A1DA76B4@ottawa.com... > I have had difficulty finding an efficient way to strip > chemiluminescent/fluorescent-detected Western blots. I recall using a > B-mercaptoethanol solution and/or an SDS solution, but I can't recall > the concentrations or the steps. Is there anyone out there who has a > protocol? > > Many thanks > Neil Use 100mM B-mercaptoethanol, 2% SDS in 62,5mM Tris (pH=6.8), at 65°C for 30min. (I used 50°C and it was still good) martin From owner-proteins@net.bio.net Sat Dec 04 03:46:38 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!Norway.EU.net!EU.net!blackbush.xlink.net!news0.de.colt.net!colt.net!newspeer.clara.net!news.clara.net!remarQ-uK!rQdQ!supernews.com!remarQ.com!remarQ69!WReNclone!WReNphoon3.POSTED!WReN!not-for-mail From: hydrophile Subject: stereospecificity and solvation Newsgroups: bionet.molbio.proteins Message-ID: <0146ce5a.364cf621@usw-ex0101-005.remarq.com> Lines: 12 Bytes: 455 X-Originating-Host: 209.180.50.92 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here X-Wren-Trace: eNz50dDJjsSPj9zG3JXN3crvwNjU2pjL1t7f1tvNi5LWztyPgsWHg4iNloyC0paZ Date: Fri, 03 Dec 1999 19:30:25 -0800 NNTP-Posting-Host: 10.0.2.5 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon3 944278417 10.0.2.5 (Fri, 03 Dec 1999 19:33:37 PST) NNTP-Posting-Date: Fri, 03 Dec 1999 19:33:37 PST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:474 Im a physical chemist transitioning to a biophysicist and would appreciate some help with a question. I know proteins exhibit stereospecific binding, but are they also stereospecific solutes? i.e. has anyone documented a protein being more soluble in one solvent isomer versus another. thanks. * Sent from RemarQ http://www.remarq.com The Internet's Discussion Network * The fastest and easiest way to search and participate in Usenet - Free! From owner-proteins@net.bio.net Sat Dec 04 22:04:35 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.easynews.net!newsfeed.germany.net!newsfeed.tli.de!news-raspail.gip.net!news.gsl.net!gip.net!news5-gui.server.ntli.net!ntli.net!not-for-mail From: Lee Hunt Newsgroups: bionet.molbio.proteins Subject: Reverse complement Date: Sat, 04 Dec 1999 22:07:27 +0000 Organization: Virgin Net Usenet Service Lines: 4 Message-ID: <3849909F.D0DF7188@virgin.net> NNTP-Posting-Host: p-33-virgin1.tch.virgin.net Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: nclient15-gui.server.virgin.net 944345071 28789 212.250.194.33 (4 Dec 1999 22:04:31 GMT) X-Complaints-To: abuse@virgin.net NNTP-Posting-Date: 4 Dec 1999 22:04:31 GMT X-Mailer: Mozilla 4.6 [en] (Win95; I) X-Accept-Language: en Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:475 Can anyone point me to any web sites that will give me the reverse complement of a genomic sequence lee From owner-proteins@net.bio.net Sun Dec 05 19:22:51 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!uni-berlin.de!pc207-60.biochem.uni-potsdam.DE!not-for-mail From: Frank Fuerst Newsgroups: bionet.molbio.proteins,bionet.biophysics Subject: Re: stereospecificity and solvation Followup-To: bionet.biophysics Date: Sun, 05 Dec 1999 20:26:43 +0100 Organization: =?iso-8859-1?Q?Universit=E4t?= Potsdam Lines: 37 Message-ID: <82ee28$2ltpf$1@fu-berlin.de> References: <0146ce5a.364cf621@usw-ex0101-005.remarq.com> NNTP-Posting-Host: pc207-60.biochem.uni-potsdam.de (141.89.207.60) Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Trace: fu-berlin.de 944421768 2815791 141.89.207.60 (16 49 955) X-Mailer: Mozilla 4.51 [de]C-CCK-MCD QXW03200 (WinNT; I) X-Accept-Language: de,en Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:476 bionet.biophysics:244 hydrophile wrote in bionet.molbio.proteins: > I know proteins exhibit stereospecific binding, but are they also > stereospecific solutes? i.e. has anyone documented a protein being more > soluble in one solvent isomer versus another. A, that's what you wanted! Well, first you would need a chiral or stereoisomeric liquid in that proteins are soluble at all. Next, if this is a hydrophobic solvent, you'll probably get denatured proteins, and denatured proteins in organic solvent is not the most relevant system, is it? Second, if you find some solvent which retains the proteins' native structure, you'll find that though the building blocks are chiral and of course the whole globular structure is often chiral, the surface consists rather randomly of backbone atoms and the more hydrophilic bunch of side chains, each in different possible rotational configuration. Thus, averaging over the whole protein surface, you wouldn't expect the protein to specifically interact with stereoisomers. Thus I think your only chance is to design something big enough to interact with the whole protein as one chiral entity, which has a size of several tens or even hundreds of Angströms - is that a solvent? Or you can build something that specifically interacts with some characteristic form on the protein surface, with a binding pocket or similar forms - but then it's not solvation, rather specific binding. X-Posted to bionet.biophysics where you posted your longer question, and Follow-up-to set. Frank -- Hi! I'm Norton Antivirus. Replace your signature with this text for protection against Signature Virus 99 and many others. From owner-proteins@net.bio.net Sun Dec 05 21:41:59 1999 Path: hgmp.mrc.ac.uk!biosci From: 186uK6Dnz@fnmail.com Newsgroups: bionet.molbio.proteins Subject: GET EXPOSED WITH BULK E-MAIL!!****HOLIDAY SPECIAL*** Date: 5 Dec 1999 21:41:59 -0000 Organization: MRC Human Genome Mapping Project Resource Centre Lines: 68 Message-ID: <199912052137.PAA23167@pioneer.digivis.com> NNTP-Posting-Host: mercury.hgmp.mrc.ac.uk X-Trace: niobium.hgmp.mrc.ac.uk 944430119 27905 193.62.192.80 (5 Dec 1999 21:41:59 GMT) X-Complaints-To: news@hgmp.mrc.ac.uk NNTP-Posting-Date: 5 Dec 1999 21:41:59 GMT X-Received: from mserv3.dl.ac.uk (IDENT:root@mserv3.dl.ac.uk [148.79.80.28]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id VAA23522 for ; Sun, 5 Dec 1999 21:41:57 GMT X-Received: from pioneer.digivis.com (pioneer.digivis.com [156.46.133.10]) by mserv3.dl.ac.uk (8.9.3/8.9.3/[ref postmaster@dl.ac.uk]) with ESMTP id VAA04689 for ; Sun, 5 Dec 1999 21:42:22 GMT X-Received: from 2PGI8cUrj by pioneer.digivis.com (8.8.7/DV Multi-Homed SMTP 3.1) id PAA23167; Sun, 5 Dec 1999 15:37:52 -0600 X-Reply-to: wecanhelp@fnmail.com X-TO: wecanhelp@fnmail.com Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:477 SORRY IF THIS AD HAS OFFENDED YOU IN ANYWAY. 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FOR ALL OTHER AMOUNTS YOU MUST CALL FOR OUR PRICING. We also offer larger list packages. You will need to call for pricing. CALL NOW 209-669-3628, WE ARE ON THE WEST COAST SO PLEASE TAKE THIS INTO CONSIDERATION WHEN CALLING. YOU MAY CALL 7 DAYS A WEEK, ANYTIME! Once you are ready to order, just e-mail or call and we will e-mail you a work order. Bulk e-mailing sells itself, so when you are ready to make a positive change in your business just let us know. Just as you responded to my e-mail ad you can have people respond to yours, so the answer to the question - "Does bulk e-mailing really work?" Yes it does. You responded to a bulk e-mail ad didn't you? :) CALL 209-669-3628 METHOD OF PAYMENT UNTIL THE HOLIDAYS ARE OVER IS, CASHIERS CHECK MONEY ORDER OR BANK WIRE. removeall@china.com to be removed --- From owner-proteins@net.bio.net Mon Dec 06 04:55:19 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news.algonet.se!algonet!newsfeed.berkeley.edu!newsgate.cuhk.edu.hk!hkusud.hku.hk!hkunae.hku.hk!hkusub.hku.hk!not-for-mail From: Xia Xuhua Newsgroups: bionet.molbio.proteins Subject: Re: Reverse complement Date: 6 Dec 1999 04:39:40 GMT Organization: The University of Hong Kong Lines: 39 Message-ID: <82femc$19he4@hkunae.hku.hk> References: <3849909F.D0DF7188@virgin.net> NNTP-Posting-Host: hkusub.hku.hk X-Newsreader: TIN [UNIX 1.3 release 961025] Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:478 Lee Hunt wrote: : Can anyone point me to any web sites that will give me the reverse : complement of a genomic sequence : lee If you have installed my program DAMBE, you can just read the sequences and click "Sequences|Get complement sequences". I have released a new version of DAMBE with many enhancement and a comprehensive help system. The software is available from my web site: http://web.hku.hk/~xxia/software/software.htm DAMBE is an integrated software package for retrieving, organizing, manipulating, aligning, graphically displaying, and analyzing molecular sequence data. Many tools for comparative sequences analysis and extensive tests of alternative phylogenetic hypotheses have been implemented. Although DAMBE is mainly for molecular sequence data, allele frequency data can also be used by DAMBE for calculating genetic distances or phylogenetic reconstruction. DAMBE runs on Windows 95/98/NT (including the MACs that installed the virtual PC software). Two installation problems have been addressed by Microsoft and I have included the links to the Microsoft solutions related to DAMBE installation. Best. Xuhua ================================================================== Dr. Xuhua Xia | Tel: (852) 2857 8239 (lab) Dept. Ecol. & Biod. | Tel: (852) 2975 5629 (office) Rm 603, T.T. Tsui Building | Fax: (852) 2517 6082 The University of Hong Kong | Email: xxia@hkusua.hku.hk Pukfulam Road | WWW: http://web.hku.hk/~xxia Hong Kong | ================================================================== From owner-proteins@net.bio.net Mon Dec 06 14:55:27 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!vixen.cso.uiuc.edu!not-for-mail From: "Rishi Shukla" Newsgroups: bionet.molbio.proteins Subject: Solubility of proteins in non aqueous solvents Date: Mon, 6 Dec 1999 08:55:53 -0600 Organization: University of Illinois at Urbana-Champaign Lines: 27 Message-ID: <82gior$jtj$1@vixen.cso.uiuc.edu> NNTP-Posting-Host: rochester-76.slip.uiuc.edu X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:479 Folks, Most proteins are soluble in water but some like insulin and zein (from corn) are predominantly soluble in nonaqueous solvents like ethanol solutions. My question is: Are there ways to predict what nonaqueous solvents or nonaqueous solutions (with water) would a protein be soluble in? I know of some work in the 60's where people just dissolved the protein in large number of them and check for solubility. But this cannot be done for all the solvents. Please don't waste your resources on directing me to the library. I am doing my searches. This post is directed towards people who may have some specific references. thank you, -r -- Rishi Shukla University of Illinois, Urbana www.ews.uiuc.edu/~rshukla -- -- Rishi Shukla University of Illinois, Urbana www.ews.uiuc.edu/~rshukla From owner-proteins@net.bio.net Tue Dec 07 10:20:03 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!newsxfer.itd.umich.edu!news.join.ad.jp!news-sv.sinet!news.nc.u-tokyo.ac.jp!gray.ims.u-tokyo.ac.jp!not-for-mail From: "Tatsuya Akutsu" Newsgroups: bionet.molbio.proteins Subject: RECOMB 2000: CALL FOR POSTERS Date: Fri, 3 Dec 1999 11:55:49 +0900 Organization: Human Genome Center, Inst. of Medical Science, Univ. of Tokyo, Japan. Lines: 39 Message-ID: <827b5c$lt5$1@gray.ims.u-tokyo.ac.jp> NNTP-Posting-Host: 133.103.74.209 Mime-Version: 1.0 Content-Type: text/plain; charset="iso-2022-jp" Content-Transfer-Encoding: 7bit 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 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:482 ****************************** CALL FOR RECOMB 2000 POSTERS ****************************** +----------------------------------------------------+ | DEADLINE: January 10, 2000 | | NOTIFICATION: January 29, 2000 | | SUBMISSION: recomb2000-poster@ims.u-tokyo.ac.jp | | URL: http://recomb2000.ims.u-tokyo.ac.jp/ | +----------------------------------------------------+ The Fourth Annual International Conference on Computational Molecular Biology (RECOMB 2000) announces a call for posters. The conference will take place in Tokyo on April 8-11, 2000. POSTER ABSTRACTS SHOULD BE SUBMITTED BY FOLLOWING EXACTLY THE +----------------------------------------------------+ | INSTRUCTIONS FOR AUTHORS | | http://recomb2000.ims.u-tokyo.ac.jp/ | | instructions_for_authors.html | +----------------------------------------------------+ Accepted posters will appear in a hard cover book titled +----------------------------------------------------+ | "Currents in Computational Molecular Biology" | | S. Miyano, R. Shamir, T. Takagi (eds.) | | Universal Academy Press, Inc. | | (http://www.uap.co.jp/) | +----------------------------------------------------+ For more information about the conference and poster session visit the web site. * We are sorry if you received this email in multiple. * If you will be removed from the RECOMB email list, please let us know (recomb2000@ims.u-tokyo.ac.jp). Thank you. From owner-proteins@net.bio.net Tue Dec 07 17:31:20 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.maxwell.syr.edu!newsfeed.tli.de!news-raspail.gip.net!news.gsl.net!gip.net!news5-gui.server.ntli.net!ntli.net!server4.netnews.ja.net!willow.cc.kcl.ac.uk!not-for-mail From: Will Fuller Newsgroups: bionet.molbio.proteins Subject: ief gels Date: Tue, 07 Dec 1999 17:30:29 +0000 Organization: Cardiovascular Research, The King's Centre for Cardiovascular Biology and Medicine, The Rayne Institute, St. Thomas's Hospital Lines: 41 Message-ID: <384D4434.F302D346@kcl.ac.uk> Reply-To: will.fuller@kcl.ac.uk NNTP-Posting-Host: rayne-175.rayne.umds.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: willow.cc.kcl.ac.uk 944587827 29455 159.92.136.175 (7 Dec 1999 17:30:27 GMT) X-Complaints-To: abuse@news.kcl.ac.uk NNTP-Posting-Date: 7 Dec 1999 17:30:27 GMT X-Mailer: Mozilla 4.7 (Macintosh; I; PPC) X-Accept-Language: en Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:483 Dear all, I am having considerable difficulty with separation of proteins on ief gels. Specifically, I am looking for a 100kDa integral membrane protein and a change in its phosphorylation state. Detection is by western analysis. I have tried all combinations of the following additions to increase the solubility of the protein in the gel and the loading buffer: 2% Triton X-100 2% NP-40 2% CHAPS 9.5M urea 5% B-mercaptoethanol Best results in terms of intensity of bands on a stained gel are with a gel consisting of 4% acrylamide (37.5:1) and 5% ampholyte with the above additions less BME. Sample buffer is the same without acrylamide but including ampholytes and BME. I focus in mini-gels at 500V for 4 hours. I can see plenty of bands on a stained ief gel and on a stained PVDF membrane after semi-dry transfer in 0.7% acetic acid. If I run the same samples on SDS PAGE I have no problem getting a signal when blotting for this membrane protein when the same amount of protein is loaded. If I blot my ief gels for soluble cytoplasmic proteins I get good signals, and I observe bands shifting on phosphorylation of these proteins. I am assuming that my membrane protein (and maybe others, I haven't looked) is not entering the gel. I have run ief gels with pH ranges from 4-6 and 3-10. The theoretical pI (not necessarily worth much, I know) of my protein is 5.1, I am fairly confident that the pH is such that it would enter the gel if it could. Does anyone out there have any thoughts or comments on how I can improve entry of membrane proteins into ief gels? Thanks! Will From owner-proteins@net.bio.net Tue Dec 07 17:58:42 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newshub.northeast.verio.net!verio!sunqbc.risq.qc.ca!carnaval.risq.qc.ca.POSTED!not-for-mail From: "Younes Bounhar" Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins References: <3846E641.A1DA76B4@ottawa.com> Subject: Re: Western stripping Lines: 19 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, 07 Dec 1999 17:58:37 GMT NNTP-Posting-Host: 198.168.191.32 X-Complaints-To: abuse@mcgill.ca X-Trace: carnaval.risq.qc.ca 944589517 198.168.191.32 (Tue, 07 Dec 1999 12:58:37 EST) NNTP-Posting-Date: Tue, 07 Dec 1999 12:58:37 EST Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3419 bionet.molbio.proteins:484 you can also use 0.2 M NaOH for 5' at RT, it works just great. (don't leave it any longer it will strip your antigen away too!!) Younes Neil McKenna wrote in message news:3846E641.A1DA76B4@ottawa.com... > I have had difficulty finding an efficient way to strip > chemiluminescent/fluorescent-detected Western blots. I recall using a > B-mercaptoethanol solution and/or an SDS solution, but I can't recall > the concentrations or the steps. Is there anyone out there who has a > protocol? > > Many thanks > Neil From owner-proteins@net.bio.net Thu Dec 09 12:10:20 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!server5.netnews.ja.net!nntp.news.xara.net!xara.net!gxn.net!baron.netcom.net.uk!netcom.net.uk!server3.netnews.ja.net!news.ox.ac.uk!not-for-mail From: =?iso-8859-1?Q?Jes=FAs?= Zurdo Alaguero Newsgroups: bionet.molbio.proteins Subject: pH-like effect of Calcium on acidic residues Date: Thu, 09 Dec 1999 12:09:57 +0000 Organization: OCMS - University of Oxford Lines: 27 Message-ID: <384F9C15.8F675299@bioch.ox.ac.uk> Reply-To: zurdo@bioch.ox.ac.uk NNTP-Posting-Host: chuchi.ocms.ox.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: news.ox.ac.uk 944741401 16049 163.1.35.189 (9 Dec 1999 12:10:01 GMT) X-Complaints-To: newsmaster@ox.ac.uk NNTP-Posting-Date: 9 Dec 1999 12:10:01 GMT X-Mailer: Mozilla 4.7 (Macintosh; I; PPC) X-Accept-Language: en,es Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:485 Dear readers I have heard about people experiencing effects similar to protonation (low pH) on acidic residues when calcium was added to protein solutions or peptides when studying folding and dynamics of those. Unfortunately I've got stuck searching the literature and I cannot find anything but pure Ca2+ interaction with calcium binding proteins, and I would be interested in a less specific effect. Does anybody know about this, and/or can provide me with any reference, name or whatsoever? Thanks very very much J.Zurdo ____________________________________________________ J. Zurdo New Chemistry Laboratory, OCMS-University of Oxford South Parks Road, Oxford OX1 3QT United Kingdom email: zurdo@bioch.ox.ac.uk Do not add this address to any database ____________________________________________________ From owner-proteins@net.bio.net Thu Dec 09 12:14:18 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!news.uhi.ac.uk!server6.netnews.ja.net!nntp.news.xara.net!xara.net!gxn.net!news.maxwell.syr.edu!newsfeed.berkeley.edu!newsfeed.stanford.edu!biosci!pharm.sunysb.edu!connelly From: connelly@pharm.sunysb.edu ("connelly") Newsgroups: bionet.molbio.proteins Subject: antibody recognition of eiptope tags Date: 8 Dec 1999 08:45:47 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 46 Sender: daemon@net.bio.net Distribution: world Message-ID: <384E89F4.F69B4B98@pharm.sunysb.edu> NNTP-Posting-Host: net.bio.net Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:486 This is a multi-part message in MIME format. --------------95312A43252278CCF7E2B26B Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit I have strategically placed myc epitopes into my protein of interest and have found that, while the site is there by sequence, only some of the sites are accessable to the monclonal antibody I am using to recognize the epitope on a Western blot. The antibody does, however, recognize the site by immunofluorescence and flow cytometry. Since I am working with a membrane protein it is possible that it is not completely denatured under the conditions I have used for electrophoresis. I have tried excess SDS, 100 mM Dtt, 8 M Urea, incubations at 37oC or boiling (in different combinations) with no success. I did get partial success in denaturing the protein further by alkylating with iodoacetamide after SDS and DTT treatment. I would like to try Guanidine HCL but have not found a protocol that would be easy to use with several samples that would need to be run by PAGE. Does anyone out there have any suggestions? Marge --------------95312A43252278CCF7E2B26B Content-Type: text/x-vcard; charset=us-ascii; name="connelly.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Margery A. Connelly Content-Disposition: attachment; filename="connelly.vcf" begin:vcard n:Connelly;Margery A. x-mozilla-html:FALSE org:SUNY at Stony Brook;Dept. of Pharmacological Sciences adr:;;BHS T7-141;Stony Brook;New York;11794-8651; version:2.1 email;internet:connelly@pharm.sunysb.edu title:Research Assistant Professor x-mozilla-cpt:;-24672 fn:Margery A. Connelly end:vcard --------------95312A43252278CCF7E2B26B-- From owner-proteins@net.bio.net Thu Dec 09 12:19:47 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!news.uhi.ac.uk!server6.netnews.ja.net!server4.netnews.ja.net!news5-gui.server.ntli.net!ntli.net!news-lond.gip.net!news.gsl.net!gip.net!newsfeed.icl.net!newsfeed.nacamar.de!sunqbc.risq.qc.ca!carnaval.risq.qc.ca.POSTED!not-for-mail Message-ID: <384E909B.313588B2@crchul.ulaval.ca> From: Alex Blais Organization: Centre de Recherche du CHUL(=?iso-8859-1?Q?Qu=E9bec?=) X-Mailer: Mozilla 4.5 [en] (WinNT; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: GST dimers Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit Lines: 23 Date: Wed, 08 Dec 1999 17:15:12 GMT NNTP-Posting-Host: 132.203.35.36 X-Complaints-To: abuse@ulaval.ca X-Trace: carnaval.risq.qc.ca 944673312 132.203.35.36 (Wed, 08 Dec 1999 12:15:12 EST) NNTP-Posting-Date: Wed, 08 Dec 1999 12:15:12 EST Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3440 bionet.molbio.proteins:487 Hi, I am trying to purify a recombinant protein fused to GST overproduced in E.coli. I have some problems during the purification steps, and I hypothesised that homodimers formation mediated by the GST moiety could be the cause of the problem. Does anybody know what conditions could be used to minimize the formation of such dimers ? I already know that there are 4 cysteines in the gst monomer and I know that they are not involved in dimer formation, since mutation to serines still allows dimer formation (these are results published by other groups). I also know that in 2 M urea, the dimers are still allowed (this is my own experience). Thanks a lot for any suggestions Alexandre Blais Graduate Student Centre de Recherche du CHUL Québec, Canada From owner-proteins@net.bio.net Thu Dec 09 12:20:08 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!server5.netnews.ja.net!nntp.news.xara.net!xara.net!gxn.net!newsfeed.icl.net!newsfeed.nacamar.de!rz.uni-karlsruhe.de!fu-berlin.de!uni-berlin.de!pc207-60.biochem.uni-potsdam.DE!not-for-mail From: Frank Fuerst Newsgroups: bionet.molbio.proteins Subject: Re: pH-like effect of Calcium on acidic residues Date: Thu, 09 Dec 1999 13:20:28 +0100 Organization: =?iso-8859-1?Q?Universit=E4t?= Potsdam Lines: 17 Message-ID: <82o6ih$2th0a$1@fu-berlin.de> References: <384F9C15.8F675299@bioch.ox.ac.uk> NNTP-Posting-Host: pc207-60.biochem.uni-potsdam.de (141.89.207.60) Mime-Version: 1.0 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: 8bit X-Trace: fu-berlin.de 944741778 3064842 141.89.207.60 (16 49 955) X-Mailer: Mozilla 4.51 [de]C-CCK-MCD QXW03200 (WinNT; I) X-Accept-Language: de,en Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:488 Jesús Zurdo Alaguero wrote: > I have heard about people experiencing effects similar to protonation > (low pH) on acidic residues when calcium was added to protein solutions > or peptides when studying folding and dynamics of those. Unfortunately > I've got stuck searching the literature and I cannot find anything but > pure Ca2+ interaction with calcium binding proteins, and I would be > interested in a less specific effect. Of course Calcium2+, as any ion, would increase the ionic strength of the solution, and this in turn alters (decreases) the effect of electrostatic interactions, e.g. salt bridges. Frank -- Hi! I'm Norton Antivirus. Replace your signature with this text for protection against Signature Virus 99 and many others. From owner-proteins@net.bio.net Thu Dec 09 15:46:55 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!nntp.cs.ubc.ca!cyclone.bc.net!clarke.sasknet.sk.ca!tribune.usask.ca!not-for-mail From: Colin Rasmussen Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: GST dimers Date: Thu, 09 Dec 1999 09:46:01 -0600 Organization: University of Saskatchewan Lines: 19 Message-ID: <384FCEB8.C0DD0FAF@pombe.usask.ca> References: <384E909B.313588B2@crchul.ulaval.ca> NNTP-Posting-Host: schizo.usask.ca Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii; x-mac-type="54455854"; x-mac-creator="4D4F5353" Content-Transfer-Encoding: 7bit X-Trace: tribune.usask.ca 944754367 12707 128.233.77.115 (9 Dec 1999 15:46:07 GMT) X-Complaints-To: usenet@tribune.usask.ca NNTP-Posting-Date: 9 Dec 1999 15:46:07 GMT X-Mailer: Mozilla 4.61 (Macintosh; U; PPC) X-Accept-Language: en Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3467 bionet.molbio.proteins:490 Alex Blais wrote: > Hi, > > I am trying to purify a recombinant protein fused to GST overproduced in > E.coli. I have some problems during the purification steps, and I > hypothesised that homodimers formation mediated by the GST moiety could > be the cause of the problem. Two questions: How do you know there are dimers? Why does that pose a problem? Colin From owner-proteins@net.bio.net Thu Dec 09 17:20:31 1999 Path: hgmp.mrc.ac.uk!guardian!warwick!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!wn4feed!worldnet.att.net!wnmaster1!not-for-mail From: "Stephen Dahl" Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: GST dimers Date: Thu, 9 Dec 1999 12:21:40 -0500 Organization: AT&T WorldNet Services Lines: 18 Message-ID: <82ooa2$hn3$1@bgtnsc03.worldnet.att.net> References: <384E909B.313588B2@crchul.ulaval.ca> NNTP-Posting-Host: 12.78.177.11 X-Trace: bgtnsc03.worldnet.att.net 944759938 18147 12.78.177.11 (9 Dec 1999 17:18:58 GMT) X-Complaints-To: abuse@worldnet.att.net NNTP-Posting-Date: 9 Dec 1999 17:18:58 GMT X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2615.200 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3468 bionet.molbio.proteins:491 Alex Blais wrote in message news:384E909B.313588B2@crchul.ulaval.ca... > Does anybody know what conditions could be used to minimize the > formation of such dimers ? Is it Amersham who sells the pGEX vector commercially now? I think it is. Anyhow, my point is that a few years ago I went to a seminar on GST provided by whichever company was marketing it and I think they mentioned a method. Unfortunately, I don't remember what it was (sorry), but a check on their website or a call to tech. service may be useful. Good luck, Steve Dahl JHMI From owner-proteins@net.bio.net Fri Dec 10 01:51:05 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!news1.tor.metronet.ca!newsfeed2.golden.net!not-for-mail From: "Attasak Jaree" Newsgroups: bionet.molbio.proteins Subject: ALBUMIN? Date: Thu, 9 Dec 1999 20:46:25 -0600 Organization: Golden Triangle Online Lines: 8 Message-ID: <82plvi$md2$1@cougar.golden.net> NNTP-Posting-Host: as52-22-89.cas-kit.golden.net X-Trace: cougar.golden.net 944790322 22946 209.183.132.89 (10 Dec 1999 01:45:22 GMT) X-Complaints-To: abuse@golden.net NNTP-Posting-Date: 10 Dec 1999 01:45:22 GMT X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2314.1300 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2314.1300 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:492 Hello, Could anyone please tell me where I can find the diffusion coefficient of Albumin in water at 297K? Thank you for your help. Attasak. From owner-proteins@net.bio.net Fri Dec 10 08:32:12 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.berkeley.edu!news.ucdavis.edu!logan.ucdavis.edu!kcweiss From: Kathryn Weiss Newsgroups: bionet.molbio.proteins Subject: o-acetyl-DL-homorserine source? Date: Thu, 9 Dec 1999 19:03:45 -0800 Organization: University of California, Davis Lines: 11 Message-ID: NNTP-Posting-Host: logan.ucdavis.edu Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Sender: kcweiss@logan.ucdavis.edu Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:493 I'm looking for a commercial source for O-acetyl-DL-homoserine. Any suggestions? I've checked the usual places, but it seems to be too exotic for them. Thanks, Kathryn Weiss Viticulture & Enology University of California, Davis From owner-proteins@net.bio.net Fri Dec 10 21:20:25 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!newsfeed.cwix.com!news.binc.net!nntp.chorus.net!news.itis.com!news.doit.wisc.edu!default From: klenchin@facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.proteins Subject: Re: GST dimers Date: Fri, 10 Dec 1999 21:18:12 GMT Organization: UW-Madison Lines: 23 Message-ID: <82rqf9$e5m$2@news.doit.wisc.edu> References: <384E909B.313588B2@crchul.ulaval.ca> NNTP-Posting-Host: martinlab.biochem.wisc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=KOI8-R Content-Transfer-Encoding: 8bit X-Newsreader: News Xpress 2.01 X-No-Archive: yes Xref: hgmp.mrc.ac.uk bionet.molbio.methds-reagnts:3490 bionet.molbio.proteins:494 :Hi, : :I am trying to purify a recombinant protein fused to GST overproduced in :E.coli. I have some problems during the purification steps, and I :hypothesised that homodimers formation mediated by the GST moiety could :be the cause of the problem. : :Does anybody know what conditions could be used to minimize the :formation of such dimers ? : :I already know that there are 4 cysteines in the gst monomer and I know :that they are not involved in dimer formation, since mutation to serines :still allows dimer formation (these are results published by other :groups). I also know that in 2 M urea, the dimers are still allowed :(this is my own experience). : From what I understand, sometimes bacterial GST forms dimers and sometimes does not. This appears to be dependent on fusion sequences, and I have not seen a whole lot you can do to modify this. I would simply switch to something else - like maltose binding protein. - Dima From owner-proteins@net.bio.net Fri Dec 10 21:20:27 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.maxwell.syr.edu!newsspool.doit.wisc.edu!news.doit.wisc.edu!default From: klenchin@facstaff.REMOVE_TO_REPLY.wisc.edu (Dima Klenchin) Newsgroups: bionet.molbio.proteins Subject: Re: pH-like effect of Calcium on acidic residues Date: Fri, 10 Dec 1999 21:15:38 GMT Organization: UW-Madison Lines: 18 Message-ID: <82rqag$e5m$1@news.doit.wisc.edu> References: <384F9C15.8F675299@bioch.ox.ac.uk> NNTP-Posting-Host: martinlab.biochem.wisc.edu Mime-Version: 1.0 Content-Type: text/plain; charset=KOI8-R Content-Transfer-Encoding: 8bit X-Newsreader: News Xpress 2.01 X-No-Archive: yes Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:495 :Dear readers :I have heard about people experiencing effects similar to protonation :(low pH) on acidic residues when calcium was added to protein solutions :or peptides when studying folding and dynamics of those. Unfortunately :I've got stuck searching the literature and I cannot find anything but :pure Ca2+ interaction with calcium binding proteins, and I would be :interested in a less specific effect. :Does anybody know about this, and/or can provide me with any reference, :name or whatsoever? : The only thing tangentially related I can recall is a simple fact that a lot of people forget that when Ca2+ is added to EGTA, the result is a strong acidification, even in buffered solutions. Thus, the world is full of artefacts where people study pH effects thinking they are Ca2+ effects. - Dima From owner-proteins@net.bio.net Sat Dec 11 09:10:11 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.maxwell.syr.edu!newsfeed.icl.net!oleane!wanadoo.fr!not-for-mail From: "Mikaël" Newsgroups: bionet.molbio.proteins Subject: arginine + ninhydrine, what colour? Date: Sat, 11 Dec 1999 10:07:17 +0100 Organization: Wanadoo, l'internet avec France Telecom Lines: 3 Message-ID: <82t4db$1ns$1@wanadoo.fr> NNTP-Posting-Host: integ-ste-genevieve-1-2-72.abo.wanadoo.fr X-Trace: wanadoo.fr 944903403 1788 193.250.166.72 (11 Dec 1999 09:10:03 GMT) X-Complaints-To: abuse@wanadoo.fr NNTP-Posting-Date: 11 Dec 1999 09:10:03 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 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:496 what is yhe color of arginine with ninhydrine? From owner-proteins@net.bio.net Sat Dec 11 23:13:03 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!logbridge.uoregon.edu!howland.erols.net!portc02.blue.aol.com!audrey04.news.aol.com!not-for-mail From: petulant88@aol.com (Petulant88) Newsgroups: bionet.molbio.proteins Subject: Re: Immunoprecipitation question Lines: 14 NNTP-Posting-Host: ladder07.news.aol.com X-Admin: news@aol.com Date: 11 Dec 1999 23:12:57 GMT References: <824mgc$154c$1@nntp4.u.washington.edu> Organization: AOL http://www.aol.com Message-ID: <19991211181257.20405.00000113@ng-fh1.aol.com> Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:498 >I am planning to do something related to immunopricipitation. I am asking >that whether I should frezze the tissue samples by liquid nitrogen >immediately after they are taken or We always freeze in liquid nitrogen first, but I don't remember why...lol. It just gets the tissue to a stable temperature faster. Xandra I make money on MEDLINE: http://www.alladvantage.com/home.asp?refid=EII195 From owner-proteins@net.bio.net Sun Dec 12 11:31:40 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.uni-stuttgart.de!news.urz.uni-heidelberg.de!not-for-mail From: Christof Gaenzler Newsgroups: bionet.molbio.proteins Subject: Re: GST dimers Date: Sun, 12 Dec 1999 12:06:30 +0000 Organization: Deutsches Krebsforschungszentrum Heidelberg Lines: 28 Message-ID: <38538FC6.49DAB554@dkfz-heidelberg.de> References: <384E909B.313588B2@crchul.ulaval.ca> <384FCEB8.C0DD0FAF@pombe.usask.ca> NNTP-Posting-Host: ppp114.urz.uni-heidelberg.de Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: news.urz.uni-heidelberg.de 944996774 16719 129.206.202.154 (12 Dec 1999 11:06:14 GMT) X-Complaints-To: usenet@news.urz.uni-heidelberg.de NNTP-Posting-Date: 12 Dec 1999 11:06:14 GMT X-Mailer: Mozilla 4.08 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:499 Hi, Colin Rasmussen wrote: > Alex Blais wrote: > > I am trying to purify a recombinant protein fused to GST overproduced in > > E.coli. I have some problems during the purification steps, and I > > hypothesised that homodimers formation mediated by the GST moiety could > > be the cause of the problem. > > Two questions: > > How do you know there are dimers? > > Why does that pose a problem? In a SDS-page, the dimers are no longer stable and one can see C-terminal degradation products that are as well purified because of the intact N-terminal GST-part. If you want to have a pure full length protein you have to do a second purification step. -- German Cancer Research Center (DKFZ) Applied Tumorvirology - ATV F0200 Christof Gaenzler INF 242 69120 Heidelberg T: +49-6221-42-4937 F: +49-6221-42-4932 From owner-proteins@net.bio.net Sun Dec 12 23:27:27 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.mathworks.com!news-out.cwix.com!newsfeed.cwix.com!news.xnet.com!not-for-mail From: "River Mouse" Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Essential amino acid requirements Date: Sun, 12 Dec 1999 17:26:34 -0600 Organization: XNet Information Systems, Inc. Lines: 20 Message-ID: <831b0q$u9$1@flood.xnet.com> NNTP-Posting-Host: gw.high-voltage.com X-Trace: flood.xnet.com 945041242 969 204.248.56.3 (12 Dec 1999 23:27:22 GMT) X-Complaints-To: abuse@xnet.com NNTP-Posting-Date: 12 Dec 1999 23:27:22 GMT 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 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:500 sci.med.nutrition:10088 misc.fitness.weights:51108 I'm reading that without an adequate balance of the eight essential amino acids, the body is incapable of synthesizing tissue from protein. Instead, surplus protein is reduced to simpler proteins for synthesizing non-essential amino acids and/or converted to glucose and potentially to fat. Another poster suggested that an excessive surplus could also result in the formation of ill-defined muscle with the potential to break down more easily. (Bulky versus lean muscle). What ratio are the eight essential proteins needed in? Do these ratios change in repairing muscles under extreme duress, i.e. in weightlifting? Where can I find reference for the levels of the essential amino acids contained in common foods? My particular interest is in vegan foods. From owner-proteins@net.bio.net Mon Dec 13 01:50:14 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!logbridge.uoregon.edu!newsfeed.stanford.edu!remarQ70!rQdQ!supernews.com!remarQ.com!remarQ69!WReNclone!WReNphoon1.POSTED!WReN!not-for-mail From: clou Subject: HSV envelop proteins purification Newsgroups: bionet.molbio.proteins Message-ID: <39d0b19c.dd1e67ed@usw-ex0108-062.remarq.com> Lines: 13 Bytes: 358 X-Originating-Host: 207.96.136.124 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here X-Wren-Trace: eCMGLi82cTtwYDskM0MvLzU5PCtoNmB7JS5scnpgdDp6azJxey1nf3s= Date: Sun, 12 Dec 1999 17:39:20 +1600 NNTP-Posting-Host: 10.0.2.62 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon1 945049774 10.0.2.62 (Sun, 12 Dec 1999 17:49:34 PST) NNTP-Posting-Date: Sun, 12 Dec 1999 17:49:34 PST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:501 Hi everybody, I'd like to know if someone knows how to isolate and purify the HSV envelop glycoproteins. If you know how, could you send me a protocol? Thank you in advance.. Eric Cloutier * Sent from AltaVista http://www.altavista.com Where you can also find related Web Pages, Images, Audios, Videos, News, and Shopping. Smart is Beautiful From owner-proteins@net.bio.net Mon Dec 13 10:21:24 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.uni-paderborn.de!news-han1.dfn.de!news.gwdg.de!not-for-mail From: mhartma1@gwdg.de (Mark Hartmann) Newsgroups: bionet.molbio.proteins Subject: deglycosylation Date: Mon, 13 Dec 1999 10:20:28 GMT Organization: Molecular Microbiology Lines: 5 Message-ID: <3854c754.296298684@news.gwdg.de> NNTP-Posting-Host: brauspc5.g2l.bio.uni-goettingen.de X-Trace: gwdu67.gwdg.de 945080479 69693 134.76.70.208 (13 Dec 1999 10:21:19 GMT) X-Complaints-To: news@gwdg.de NNTP-Posting-Date: 13 Dec 1999 10:21:19 GMT X-Newsreader: Forte Free Agent 1.11/32.235 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:502 Hi Everybody, has anyone a good and easy protocol to deglycosylate proteins ? Regards Mark From owner-proteins@net.bio.net Mon Dec 13 11:46:34 1999 From: "Jules" <2juls@one.net.au> Subject: Re: Essential amino acid requirements Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights References: <831b0q$u9$1@flood.xnet.com> Message-ID: <01bf455f$9a1154e0$994065cb@default> X-Newsreader: Microsoft Internet News 4.70.1162 NNTP-Posting-Host: 203.101.64.153 Date: 13 Dec 1999 22:45:35 +1000 X-Trace: 13 Dec 1999 22:45:35 +1000, 203.101.64.153 Lines: 32 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!feeder.qis.net!logbridge.uoregon.edu!newsfeed.stanford.edu!paloalto-snf1.gtei.net!news.gtei.net!newsgate.tandem.com!uunet!pao.uu.net!lax.uu.net!pink.one.net.au!203.101.64.153 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:503 sci.med.nutrition:10145 misc.fitness.weights:51217 Diet for a small planet, by Frances Moore Lappe. Go to a library, it's over 20 years old. If they don't have it ask them to get it in on inter-library loan. There may be newer books, but this is the only one I know with this much detail. jules River Mouse wrote in article <831b0q$u9$1@flood.xnet.com>... > I'm reading that without an adequate balance of the eight essential amino > acids, the body is incapable of synthesizing tissue from protein. Instead, > surplus protein is reduced to simpler proteins for synthesizing > non-essential amino acids and/or converted to glucose and potentially to > fat. > > Another poster suggested that an excessive surplus could also result in the > formation of ill-defined muscle with the potential to break down more > easily. (Bulky versus lean muscle). > > > What ratio are the eight essential proteins needed in? Do these ratios > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > Where can I find reference for the levels of the essential amino acids > contained in common foods? My particular interest is in vegan foods. > > > > From owner-proteins@net.bio.net Mon Dec 13 14:55:04 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news-feed.ifi.uio.no!howland.erols.net!outgoing.news.rcn.net.MISMATCH!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Mon, 13 Dec 1999 09:53:31 +0000 Lines: 40 Message-ID: <3854C21D.4A7D@erols.com> References: <831b0q$u9$1@flood.xnet.com> <01bf455f$9a1154e0$994065cb@default> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: w+v70YnHKQy/DwrD5I3QzC+in/tL5jAqYaxJMmM5Ke4= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 13 Dec 1999 14:51:58 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:505 sci.med.nutrition:10155 misc.fitness.weights:51246 Jules wrote: > > Diet for a small planet, by Frances Moore Lappe. Go to a library, it's > over 20 years old. If they don't have it ask them to get it in on > inter-library loan. There may be newer books, but this is the only one I > know with this much detail. > jules No, don't read diet for a small planet. One edition of that book contained inaccurate information on protein. Frances Moore Lappe claims that you need to "complete" proteins, and besides, she thinks we can solve the problem of world hunger without getting rid of socialist third world dictators. It's not meat that's the cause of the famine in Africa; it's socialism that is the source of the trouble. Meat is what makes animals suffer, not humans. > > River Mouse wrote in article > <831b0q$u9$1@flood.xnet.com>... > > I'm reading that without an adequate balance of the eight essential amino > > acids, the body is incapable of synthesizing tissue from protein. > Instead, > > surplus protein is reduced to simpler proteins for synthesizing > > non-essential amino acids and/or converted to glucose and potentially to > > fat. > > > > Another poster suggested that an excessive surplus could also result in > the > > formation of ill-defined muscle with the potential to break down more > > easily. (Bulky versus lean muscle). > > > > > > What ratio are the eight essential proteins needed in? Do these ratios > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > > > Where can I find reference for the levels of the essential amino acids > > contained in common foods? My particular interest is in vegan foods. > > > > > > > > From owner-proteins@net.bio.net Mon Dec 13 14:55:04 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!howland.erols.net!outgoing.news.rcn.net.MISMATCH!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Mon, 13 Dec 1999 09:49:39 +0000 Lines: 23 Message-ID: <3854C135.2428@erols.com> References: <831b0q$u9$1@flood.xnet.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: uICjBSIvnDlFFDdRHI6h6Sg62x1ENUmivZGXdR4U/XA= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 13 Dec 1999 14:48:06 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:504 sci.med.nutrition:10154 misc.fitness.weights:51245 River Mouse wrote: > > I'm reading that without an adequate balance of the eight essential amino > acids, the body is incapable of synthesizing tissue from protein. Instead, > surplus protein is reduced to simpler proteins for synthesizing > non-essential amino acids and/or converted to glucose and potentially to > fat. > > Another poster suggested that an excessive surplus could also result in the > formation of ill-defined muscle with the potential to break down more > easily. (Bulky versus lean muscle). > > What ratio are the eight essential proteins needed in? Do these ratios > change in repairing muscles under extreme duress, i.e. in weightlifting? > > Where can I find reference for the levels of the essential amino acids > contained in common foods? My particular interest is in vegan foods. It's not the ratios you need to be concerned with, it's the amounts. As a rule of thumb, anyone eating at least 45 grams of protein a day from sources other than gelatin and watermelon is safe, even if they have active lifestyles. Except for sweets and greasy foods, all commonly eaten vegan foods have "complete" proteins. Go to Walton Feed's website. It has more details. From owner-proteins@net.bio.net Mon Dec 13 15:15:06 1999 Path: hgmp.mrc.ac.uk!pegasus.csx.cam.ac.uk!daresbury!uninett.no!news.maxwell.syr.edu!newshub2.home.com!news.home.com!news2.rdc1.on.home.com.POSTED!not-for-mail From: "Mistress Krista" Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights References: <831b0q$u9$1@flood.xnet.com> <01bf455f$9a1154e0$994065cb@default> <3854C21D.4A7D@erols.com> Subject: Re: Essential amino acid requirements Lines: 30 X-Priority: 3 X-MSMail-Priority: Normal X-Newsreader: Microsoft Outlook Express 5.00.2615.200 X-MimeOLE: Produced By Microsoft MimeOLE V5.00.2615.200 Message-ID: <81854.19750$eh2.347809@news2.rdc1.on.home.com> Date: Mon, 13 Dec 1999 15:12:04 GMT NNTP-Posting-Host: 24.114.45.168 X-Complaints-To: abuse@home.net X-Trace: news2.rdc1.on.home.com 945097924 24.114.45.168 (Mon, 13 Dec 1999 07:12:04 PST) NNTP-Posting-Date: Mon, 13 Dec 1999 07:12:04 PST Organization: @Home Network Canada Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:506 sci.med.nutrition:10158 misc.fitness.weights:51252 Stuart Dunn wrote in message news:3854C21D.4A7D@erols.com... detail. > > jules > No, don't read diet for a small planet. One edition of that book > contained inaccurate information on protein. Frances Moore Lappe claims > that you need to "complete" proteins, and besides, she thinks we can > solve the problem of world hunger without getting rid of socialist third > world dictators. It's not meat that's the cause of the famine in Africa; > it's socialism that is the source of the trouble. Actually, it's civil war, the legacy of colonialism, boundary disputes, political instability, tribal and interfaith infighting, cash cropping, natural disasters, and funding of terrorist/militia/rebel/state-sanctioned thug groups by foreign investors. You can't just replace one reductive variable like "meat" with another one like "socialism" and expect to produce a decent analysis. Krista -- ------------------------- http://krista.tico.com/weights.html mistresskrista@home.com From owner-proteins@net.bio.net Mon Dec 13 16:20:54 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!nntp.abs.net!chnws02.mediaone.net!24.128.1.101!chnws05.ne.mediaone.net!24.128.44.7!wbnws01.ne.mediaone.net.POSTED!not-for-mail Message-ID: <3854C776.C5E2015A@mediaone.net> From: ResNovae Reply-To: resnovae_I_DON'T_EAT_SPAM_@SAM.I.AM.mediaone.net X-Sender: " ResNovae" (Unverified) X-Mailer: Mozilla 4.06 [en]C-gatewaynet (Win98; I) MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <01bf455f$9a1154e0$994065cb@default> <3854C21D.4A7D@erols.com> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 61 Date: Mon, 13 Dec 1999 05:16:22 -0500 NNTP-Posting-Host: 24.147.9.134 X-Complaints-To: abuse@mediaone.net X-Trace: wbnws01.ne.mediaone.net 945102044 24.147.9.134 (Mon, 13 Dec 1999 11:20:44 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 11:20:44 EST Organization: Road Runner Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:507 sci.med.nutrition:10161 misc.fitness.weights:51270 The most recent edition retracts the original's concern re: getting complete protein at every meal. Your body can break down amino acids, store them indefinately, and then recombine them into complete proteins when the other components become available, so "protein combining" at every meal isn't neccessary like the author first believed. It's still a good book, but the older editionsaredefinately outdated. Stuart Dunn wrote: > > Jules wrote: > > > > Diet for a small planet, by Frances Moore Lappe. Go to a library, it's > > over 20 years old. If they don't have it ask them to get it in on > > inter-library loan. There may be newer books, but this is the only one I > > know with this much detail. > > jules > No, don't read diet for a small planet. One edition of that book > contained inaccurate information on protein. Frances Moore Lappe claims > that you need to "complete" proteins, and besides, she thinks we can > solve the problem of world hunger without getting rid of socialist third > world dictators. It's not meat that's the cause of the famine in Africa; > it's socialism that is the source of the trouble. Meat is what makes > animals suffer, not humans. > > > > River Mouse wrote in article > > <831b0q$u9$1@flood.xnet.com>... > > > I'm reading that without an adequate balance of the eight essential amino > > > acids, the body is incapable of synthesizing tissue from protein. > > Instead, > > > surplus protein is reduced to simpler proteins for synthesizing > > > non-essential amino acids and/or converted to glucose and potentially to > > > fat. > > > > > > Another poster suggested that an excessive surplus could also result in > > the > > > formation of ill-defined muscle with the potential to break down more > > > easily. (Bulky versus lean muscle). > > > > > > > > > What ratio are the eight essential proteins needed in? Do these ratios > > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > > > > > > Where can I find reference for the levels of the essential amino acids > > > contained in common foods? My particular interest is in vegan foods. > > > > > > > > > > > > -- Catherine Caruso ccaruso@mediaone.net ICQ#: 46412288 "The difference between fiction and reality? Fiction has to make sense." -Tom Clancy From owner-proteins@net.bio.net Mon Dec 13 16:29:10 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!bignews.mediaways.net!diablo.theplanet.net!remarQ-uK!rQdQ!supernews.com!remarQ.com!remarQ69!WReNclone!WReNphoon1.POSTED!WReN!not-for-mail From: pharveng Subject: Native electrophoresis gel Newsgroups: bionet.molbio.proteins Message-ID: <1cb3e3d0.34b5e35d@usw-ex0109-066.remarq.com> Lines: 10 Bytes: 394 X-Originating-Host: 134.184.129.3 Organization: http://www.remarq.com: The World's Usenet/Discussions Start Here X-Wren-Trace: eDQROTghZixnZCg9I2IwMyEHMT0zfnUhfzo+d2xzeTpqaiB7ayZ4dGs= Date: Mon, 13 Dec 1999 09:14:37 -0800 NNTP-Posting-Host: 10.0.2.66 X-Complaints-To: wrenabuse@remarq.com X-Trace: WReNphoon1 945102485 10.0.2.66 (Mon, 13 Dec 1999 08:28:05 PST) NNTP-Posting-Date: Mon, 13 Dec 1999 08:28:05 PST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:508 I would like to find a protocole which could allow to resolve membrane proteins on acrylamide gels. The problem is that proteins of interest are soluble in Genapol only. If you have any suggestion about this, please conatct me ! * Sent from AltaVista http://www.altavista.com Where you can also find related Web Pages, Images, Audios, Videos, News, and Shopping. Smart is Beautiful From owner-proteins@net.bio.net Mon Dec 13 16:34:35 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.mathworks.com!news-out.cwix.com!newsfeed.cwix.com!news.xnet.com!not-for-mail From: "River Mouse" Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Mon, 13 Dec 1999 10:33:23 -0600 Organization: XNet Information Systems, Inc. Lines: 78 Message-ID: <833766$35e$1@flood.xnet.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> NNTP-Posting-Host: gw.high-voltage.com X-Trace: flood.xnet.com 945102854 3246 204.248.56.3 (13 Dec 1999 16:34:14 GMT) X-Complaints-To: abuse@xnet.com NNTP-Posting-Date: 13 Dec 1999 16:34:14 GMT 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 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:509 sci.med.nutrition:10163 misc.fitness.weights:51271 "Stuart Dunn" wrote in message news:3854C135.2428@erols.com... > River Mouse wrote: > > > > I'm reading that without an adequate balance of the eight essential amino > > acids, the body is incapable of synthesizing tissue from protein. Instead, > > surplus protein is reduced to simpler proteins for synthesizing > > non-essential amino acids and/or converted to glucose and potentially to > > fat. [...] > > What ratio are the eight essential proteins needed in? Do these ratios > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > Where can I find reference for the levels of the essential amino acids > > contained in common foods? My particular interest is in vegan foods. > > It's not the ratios you need to be concerned with, it's the amounts. As > a rule of thumb, anyone eating at least 45 grams of protein a day from > sources other than gelatin and watermelon is safe, even if they have > active lifestyles. Except for sweets and greasy foods, all commonly > eaten vegan foods have "complete" proteins. Go to Walton Feed's website. > It has more details. This is a grossly inaccurate oversimplification. I know for a fact that many foods are lacking in some of the 8 essential amino acids - these are the proteins which your body CANNOT synthesize on its own. Without all eight of the essential amino acids present, your body CANNOT do anything with that protein except break it down to synthesize the nonessential proteins and treat any surplus like fat. Let's say you take 100g of each of these four protein supplements (I'm listing supplements because I can't find Essential Amino Acid ratios for conventional foods yet). Whey: Cross-Flow Microfiltration Whey Protein Isolate Soy: 90% Soy Protein Isolate Egg: Egg White Protein Concentrate Rice: Rice Protein Concentrate Here's how many grams of each of these amino acids you'd get: (You might want to change to a fixed width font for this table) Whey | Soy | Egg | Rice | EAA -----+-----|-----|------|---------- 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine 10.9 | 7.2 | 6.8 | 8.6 | L-Leucine 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan 6.4 | 4.4 | 5.1 | 4.7 | L-Valine If methionine is the limiting factor, you'll need over twice as much soy protein as you'd need whey protein to get the same usable EAA. Look at lysine, and you'll see that you need about three times as much rice as you'd need whey. Some foods are heavier in sources than others. Since I want to rely largely on soy sources for protein for the time being, I'm trying to find vegan foods which are balanced in favor of lysine, methionine, threonine and tryptophan to make up the difference. Peas and lentils are supposedly heavier in these, so the combination of peas and soy protein may result in something like whey's balance. Mind you - that's -whey's- balance. I'm still looking for a source of information on what ratios a body really needs. It may turn out that tryptophan or one of the others isn't needed in as great a proportion. From owner-proteins@net.bio.net Mon Dec 13 16:58:39 1999 Path: hgmp.mrc.ac.uk!biosci From: DOCTORHIM@aol.com Newsgroups: bionet.molbio.proteins Subject: Re: deglycosylation Date: 13 Dec 1999 16:58:39 -0000 Organization: MRC Human Genome Mapping Project Resource Centre Lines: 5 Message-ID: <0.da294d52.25867f78@aol.com> NNTP-Posting-Host: mercury.hgmp.mrc.ac.uk Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit X-Trace: niobium.hgmp.mrc.ac.uk 945104319 15147 193.62.192.80 (13 Dec 1999 16:58:39 GMT) X-Complaints-To: news@hgmp.mrc.ac.uk NNTP-Posting-Date: 13 Dec 1999 16:58:39 GMT X-Received: from imo16.mx.aol.com (imo16.mx.aol.com [152.163.225.6]) by hgmp.mrc.ac.uk (8.9.3/8.9.3) with ESMTP id QAA17114 for ; Mon, 13 Dec 1999 16:58:35 GMT X-Received: from DOCTORHIM@aol.com by imo16.mx.aol.com (mail_out_v24.6.) id 4.0.da294d52 (3939); Mon, 13 Dec 1999 11:57:29 -0500 (EST) X-To: mhartma1@gwdg.de, proteins@hgmp.mrc.ac.uk X-Mailer: AOL for Macintosh sub 53 Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:510 Mark, You can read about methods in the deglycosylation kit booklet that you can download in PDF format from http://www.prozyme.com/glycopro/index.html. --- From owner-proteins@net.bio.net Mon Dec 13 18:49:04 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!vixen.cso.uiuc.edu!sbnaran From: sbnaran@uiuc.edu (Siemel B. Naran) Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: 13 Dec 1999 18:49:01 GMT Organization: University of Illinois at Urbana-Champaign Lines: 62 Message-ID: References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Reply-To: sbnaran@uiuc.edu NNTP-Posting-Host: arboria-16.slip.uiuc.edu User-Agent: slrn/0.9.5.7 (UNIX) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:511 sci.med.nutrition:10173 misc.fitness.weights:51306 On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse >Without all eight of the essential amino acids present, your body CANNOT do >anything with that protein except break it down to synthesize the >nonessential proteins and treat any surplus like fat. Well, the body may burn some muscle to make up for the difference (ie, for whatever amino acids are lacking). Then the protein you eat is fully usable. But of course, we don't want this. >Whey | Soy | Egg | Rice | EAA >-----+-----|-----|------|---------- > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine >If methionine is the limiting factor, you'll need over twice as much soy >protein as you'd need whey protein to get the same usable EAA. > >Look at lysine, and you'll see that you need about three times as much rice >as you'd need whey. If you eat twice as much soy, you get twice as much of every amino acid. Which means too much protein, and therefore lots of work on your kidneys to get rid of the excess protein/nitrogen. >Some foods are heavier in sources than others. Since I want to rely largely >on soy sources for protein for the time being, I'm trying to find vegan >foods which are balanced in favor of lysine, methionine, threonine and >tryptophan to make up the difference. Peas and lentils are supposedly >heavier in these, so the combination of peas and soy protein may result in >something like whey's balance. Grains are high in methionine, and maybe the others. So wheat cereal and soy milk is probably a good choice. I use tofu in my spaghetti (because it tastes good to me), and this might be balance too because tofu is soy and sphaghetti is grain. >Mind you - that's -whey's- balance. I'm still looking for a source of >information on what ratios a body really needs. It may turn out that >tryptophan or one of the others isn't needed in as great a proportion. I think whole egg has the balance the body needs. I looked hard on the internet for a profile but couldn't find one (all I get are links to products and info that I already know -- what the essential amino acids are and what each one supposedly does). Anyway, as whey protein has a BV of over 100 on rats, I'd say it has the right ratios. -- -------------- siemel b naran -------------- From owner-proteins@net.bio.net Mon Dec 13 18:49:04 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!vixen.cso.uiuc.edu!sbnaran From: sbnaran@uiuc.edu (Siemel B. Naran) Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: 13 Dec 1999 18:49:02 GMT Organization: University of Illinois at Urbana-Champaign Lines: 32 Message-ID: References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Reply-To: sbnaran@uiuc.edu NNTP-Posting-Host: arboria-16.slip.uiuc.edu User-Agent: slrn/0.9.5.7 (UNIX) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:512 sci.med.nutrition:10175 misc.fitness.weights:51307 On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse >Whey | Soy | Egg | Rice | EAA >-----+-----|-----|------|---------- > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine And L-Histidine is essential for children. But I'm thinking, does the body need an absolute amount of protein everyday or an absolute ratio? If your body needs a fixed amount everyday, like 2.5 grams of methionine, then you could just eats of lots of soy and you'd surely get or exceed the minimum requirement of all the proteins. But through excess, you'd overtax your kidneys in getting rid of the nitrogen. If your body needs a fixed ratio everyday, like lysine/methionine should be 3.8 even though amount of methionine may be 1.5 or 2.5 or 3.5 grams, then no amount of soy will satisfy the requirement. -- -------------- siemel b naran -------------- From owner-proteins@net.bio.net Tue Dec 14 04:11:24 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!brick.direct.ca!quark.idirect.com.POSTED!not-for-mail Message-ID: <3855C34C.F5B6CF8D@internet.look.ca> From: Tom Matthews Organization: Longevity Unlimited X-Mailer: Mozilla 4.5 [en]C-CCK-MCD LOOKNetscape (Win95; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 45 Date: Tue, 14 Dec 1999 04:11:21 GMT NNTP-Posting-Host: 216.154.88.229 X-Complaints-To: abuse@idirect.com X-Trace: quark.idirect.com 945144681 216.154.88.229 (Mon, 13 Dec 1999 23:11:21 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 23:11:21 EST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:513 sci.med.nutrition:10213 misc.fitness.weights:51434 Stuart Dunn wrote: > > River Mouse wrote: > > > > I'm reading that without an adequate balance of the eight essential amino > > acids, the body is incapable of synthesizing tissue from protein. Instead, > > surplus protein is reduced to simpler proteins for synthesizing > > non-essential amino acids and/or converted to glucose and potentially to > > fat. > > > > Another poster suggested that an excessive surplus could also result in the > > formation of ill-defined muscle with the potential to break down more > > easily. (Bulky versus lean muscle). > > > > What ratio are the eight essential proteins needed in? Do these ratios > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > Where can I find reference for the levels of the essential amino acids > > contained in common foods? My particular interest is in vegan foods. > It's not the ratios you need to be concerned with, it's the amounts. As > a rule of thumb, anyone eating at least 45 grams of protein a day from > sources other than gelatin and watermelon is safe, even if they have > active lifestyles. Except for sweets and greasy foods, all commonly > eaten vegan foods have "complete" proteins. Go to Walton Feed's website. > It has more details. Stuart, since you obviously do not know what you are talking about, would you please stop jumping in to answer questions with highly incorrect answers! This is a science newsgroup, not a rumor mill! Please lurk for a while, read some texts, and solidify and correct your knowledge for a while instead of leading innocent people astray. --Tom Tom Matthews The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 A non-profit membership organization dedicated to the extension of the healthy human lifespan through ground breaking research, innovative ideas and practical methods. LIFE EXTENSION MAGAZINE - The ultimate source for new health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 04:13:51 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!brick.direct.ca!quark.idirect.com.POSTED!not-for-mail Message-ID: <3855C3DF.1BA33C6@internet.look.ca> From: Tom Matthews Organization: Longevity Unlimited X-Mailer: Mozilla 4.5 [en]C-CCK-MCD LOOKNetscape (Win95; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 88 Date: Tue, 14 Dec 1999 04:13:48 GMT NNTP-Posting-Host: 216.154.88.229 X-Complaints-To: abuse@idirect.com X-Trace: quark.idirect.com 945144828 216.154.88.229 (Mon, 13 Dec 1999 23:13:48 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 23:13:48 EST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:514 sci.med.nutrition:10215 misc.fitness.weights:51437 You see, Stuart! Even the person asking the question knows more than you! Talk about the "blind leading the blind"!!! --Tom River Mouse wrote: > > "Stuart Dunn" wrote in message > news:3854C135.2428@erols.com... > > River Mouse wrote: > > > > > > I'm reading that without an adequate balance of the eight essential > amino > > > acids, the body is incapable of synthesizing tissue from protein. > Instead, > > > surplus protein is reduced to simpler proteins for synthesizing > > > non-essential amino acids and/or converted to glucose and potentially to > > > fat. > [...] > > > What ratio are the eight essential proteins needed in? Do these ratios > > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > > > Where can I find reference for the levels of the essential amino acids > > > contained in common foods? My particular interest is in vegan foods. > > > > It's not the ratios you need to be concerned with, it's the amounts. As > > a rule of thumb, anyone eating at least 45 grams of protein a day from > > sources other than gelatin and watermelon is safe, even if they have > > active lifestyles. Except for sweets and greasy foods, all commonly > > eaten vegan foods have "complete" proteins. Go to Walton Feed's website. > > It has more details. > > This is a grossly inaccurate oversimplification. I know for a fact that many > foods are lacking in some of the 8 essential amino acids - these are the > proteins which your body CANNOT synthesize on its own. > > Without all eight of the essential amino acids present, your body CANNOT do > anything with that protein except break it down to synthesize the > nonessential proteins and treat any surplus like fat. > > Let's say you take 100g of each of these four protein supplements (I'm > listing supplements because I can't find Essential Amino Acid ratios for > conventional foods yet). > > Whey: Cross-Flow Microfiltration Whey Protein Isolate > Soy: 90% Soy Protein Isolate > Egg: Egg White Protein Concentrate > Rice: Rice Protein Concentrate > > Here's how many grams of each of these amino acids you'd get: > > (You might want to change to a fixed width font for this table) > > Whey | Soy | Egg | Rice | EAA > -----+-----|-----|------|---------- > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > 10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > If methionine is the limiting factor, you'll need over twice as much soy > protein as you'd need whey protein to get the same usable EAA. > > Look at lysine, and you'll see that you need about three times as much rice > as you'd need whey. > > Some foods are heavier in sources than others. Since I want to rely largely > on soy sources for protein for the time being, I'm trying to find vegan > foods which are balanced in favor of lysine, methionine, threonine and > tryptophan to make up the difference. Peas and lentils are supposedly > heavier in these, so the combination of peas and soy protein may result in > something like whey's balance. > > Mind you - that's -whey's- balance. I'm still looking for a source of > information on what ratios a body really needs. It may turn out that > tryptophan or one of the others isn't needed in as great a proportion. The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 A non-profit membership organization dedicated to the extension of the healthy human lifespan through ground breaking research, innovative ideas and practical methods. LIFE EXTENSION MAGAZINE - The ultimate source for new health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 04:32:29 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!newsfeed.direct.ca!brick.direct.ca!quark.idirect.com.POSTED!not-for-mail Message-ID: <3855C83B.B018D5AD@internet.look.ca> From: Tom Matthews Organization: Longevity Unlimited X-Mailer: Mozilla 4.5 [en]C-CCK-MCD LOOKNetscape (Win95; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 103 Date: Tue, 14 Dec 1999 04:32:25 GMT NNTP-Posting-Host: 216.154.88.229 X-Complaints-To: abuse@idirect.com X-Trace: quark.idirect.com 945145945 216.154.88.229 (Mon, 13 Dec 1999 23:32:25 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 23:32:25 EST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:515 sci.med.nutrition:10216 misc.fitness.weights:51443 Am I the only one who has noticed that the level of discourse on this newsgroup (smn) is at an all-time low? "Siemel B. Naran" wrote: > > On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse > > >Without all eight of the essential amino acids present, your body CANNOT do > >anything with that protein except break it down to synthesize the > >nonessential proteins and treat any surplus like fat. > > Well, the body may burn some muscle to make up for the difference (ie, for > whatever amino acids are lacking). Then the protein you eat is fully usable. That is a really asinine answer! That is like saying that you don't need to eat at all because you can just digest your body from the inside! > But of course, we don't want this. Do you really need to say that? > >Whey | Soy | Egg | Rice | EAA > >-----+-----|-----|------|---------- > > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > >If methionine is the limiting factor, you'll need over twice as much soy > >protein as you'd need whey protein to get the same usable EAA. > > > >Look at lysine, and you'll see that you need about three times as much rice > >as you'd need whey. > > If you eat twice as much soy, you get twice as much of every amino acid. > Which means too much protein, and therefore lots of work on your kidneys > to get rid of the excess protein/nitrogen. "Lots of work" does not equal health problems or too much protein! It has never been shown that high protein diets harm healthy kidneys. Please learn some facts before answering! > >Some foods are heavier in sources than others. Since I want to rely largely > >on soy sources for protein for the time being, It is unwise to rely on single or even restricted numbers of sources of all vital nutrients. > >I'm trying to find vegan > >foods which are balanced in favor of lysine, methionine, threonine and > >tryptophan to make up the difference. Peas and lentils are supposedly > >heavier in these, so the combination of peas and soy protein may result in > >something like whey's balance. No, the sulfur containing amino acids (methionine/cysteine) will still be limiting. Grains are what you need to balance beans/legumes. > Grains are high in methionine, and maybe the others. So wheat cereal and > soy milk is probably a good choice. I use tofu in my spaghetti (because > it tastes good to me), and this might be balance too because tofu is soy > and sphaghetti is grain. Now you are talking. But is it still nowhere as good an amino acid profile as fish/meat/dairy/whole egg is. > >Mind you - that's -whey's- balance. I'm still looking for a source of > >information on what ratios a body really needs. It may turn out that > >tryptophan or one of the others isn't needed in as great a proportion. Just fine a table of the RDAs for the essential amino acids and you will have a rough indication of the ratio of body needs. Sorry, I don't know where one is on-line and I don't have time to type it in. > I think whole egg has the balance the body needs. I looked hard on > the internet for a profile but couldn't find one (all I get are links > to products and info that I already know -- what the essential amino > acids are and what each one supposedly does). Anyway, as whey protein > has a BV of over 100 on rats, I'd say it has the right ratios. Whole egg is one of the best available. Actually (as I have said before) whole human is probably best, whole monkey next and whole cow, whole chicken or whole fish not two bad. Animal muscle meat only is not that great, because we make more than simply muscle proteins. But if you are body building animal muscle meat may be best of all. --Tom Tom Matthews The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 A non-profit membership organization dedicated to the extension of the healthy human lifespan through ground breaking research, innovative ideas and practical methods. LIFE EXTENSION MAGAZINE - The ultimate source for new health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 04:38:49 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!logbridge.uoregon.edu!newsfeed.direct.ca!brick.direct.ca!quark.idirect.com.POSTED!not-for-mail Message-ID: <3855C9B8.3C62C7C3@internet.look.ca> From: Tom Matthews Organization: Longevity Unlimited X-Mailer: Mozilla 4.5 [en]C-CCK-MCD LOOKNetscape (Win95; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 33 Date: Tue, 14 Dec 1999 04:38:45 GMT NNTP-Posting-Host: 216.154.88.229 X-Complaints-To: abuse@idirect.com X-Trace: quark.idirect.com 945146325 216.154.88.229 (Mon, 13 Dec 1999 23:38:45 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 23:38:45 EST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:516 sci.med.nutrition:10217 misc.fitness.weights:51444 "Siemel B. Naran" wrote: > If your body needs a fixed amount everyday, like 2.5 grams of methionine, > then you could just eats of lots of soy and you'd surely get or exceed the > minimum requirement of all the proteins. But through excess, you'd > overtax your kidneys in getting rid of the nitrogen. Once more with feeling, READ MY LIPS! There is no evidence, that high dietary protein is harmful for normally healthy kidneys! > If your body needs a fixed ratio everyday, like lysine/methionine should > be 3.8 even though amount of methionine may be 1.5 or 2.5 or 3.5 grams, > then no amount of soy will satisfy the requirement. You body *never* needs a fixed ratio. It only needs a minimum amount and only can use them to build protein in a certain ratio (which actually varies with the type of activity and state of your body and health). Beyond the limiting protein being used up in that ratio, all the others are use for fuel, or to make neurotransmitters and other speciallized things made from some single aminos (like niacin made from tryptophan), and the rest are excreted. --Tom Tom Matthews The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 A non-profit membership organization dedicated to the extension of the healthy human lifespan through ground breaking research, innovative ideas and practical methods. LIFE EXTENSION MAGAZINE - The ultimate source for new health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 04:41:35 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.direct.ca!brick.direct.ca!quark.idirect.com.POSTED!not-for-mail Message-ID: <3855CA60.6B29D90E@internet.look.ca> From: Tom Matthews Organization: Longevity Unlimited X-Mailer: Mozilla 4.5 [en]C-CCK-MCD LOOKNetscape (Win95; I) X-Accept-Language: en MIME-Version: 1.0 Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements References: <831b0q$u9$1@flood.xnet.com> <01bf455f$9a1154e0$994065cb@default> <3854C21D.4A7D@erols.com> <3854C776.C5E2015A@mediaone.net> Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Lines: 24 Date: Tue, 14 Dec 1999 04:41:32 GMT NNTP-Posting-Host: 216.154.88.229 X-Complaints-To: abuse@idirect.com X-Trace: quark.idirect.com 945146492 216.154.88.229 (Mon, 13 Dec 1999 23:41:32 EST) NNTP-Posting-Date: Mon, 13 Dec 1999 23:41:32 EST Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:517 sci.med.nutrition:10218 misc.fitness.weights:51448 ResNovae wrote: > > The most recent edition retracts the original's concern re: getting > complete protein at every meal. > > Your body can break down amino acids, store them indefinately, and then > recombine them into complete proteins when the other components become > available, so "protein combining" at every meal isn't neccessary like > the author first believed. > > It's still a good book, but the older editionsaredefinately outdated. Forget crap like these popular books! Buy a good nutrition text and learn some *real* nutritional science! --Tom Tom Matthews The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 A non-profit membership organization dedicated to the extension of the healthy human lifespan through ground breaking research, innovative ideas and practical methods. LIFE EXTENSION MAGAZINE - The ultimate source for new health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 14:15:24 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!enews.sgi.com!newsfeed1.funet.fi!ousrvr3.oulu.fi!sun3.oulu.fi!pkursula From: Petri Kursula Newsgroups: bionet.molbio.proteins Subject: Re: GST dimers Date: Tue, 14 Dec 1999 16:10:19 +0200 Organization: University of Oulu Lines: 45 Message-ID: References: <384E909B.313588B2@crchul.ulaval.ca> <384FCEB8.C0DD0FAF@pombe.usask.ca> <38538FC6.49DAB554@dkfz-heidelberg.de> NNTP-Posting-Host: sun3.oulu.fi Mime-Version: 1.0 Content-Type: TEXT/PLAIN; charset=US-ASCII X-Trace: ousrvr3.oulu.fi 945180621 25434 130.231.240.13 (14 Dec 1999 14:10:21 GMT) X-Complaints-To: news@news.oulu.fi NNTP-Posting-Date: 14 Dec 1999 14:10:21 GMT X-Sender: pkursula@sun3.oulu.fi In-Reply-To: <38538FC6.49DAB554@dkfz-heidelberg.de> Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:518 I would start by playing with the induction conditions there...time, IPTG concentration, temperature etc. On Sun, 12 Dec 1999, Christof Gaenzler wrote: > Hi, > > Colin Rasmussen wrote: > > Alex Blais wrote: > > > I am trying to purify a recombinant protein fused to GST overproduced in > > > E.coli. I have some problems during the purification steps, and I > > > hypothesised that homodimers formation mediated by the GST moiety could > > > be the cause of the problem. > > > > Two questions: > > > > How do you know there are dimers? > > > > Why does that pose a problem? > > In a SDS-page, the dimers are no longer stable and one can see C-terminal > degradation products that are as well purified because of the intact > N-terminal GST-part. If you want to have a pure full length protein you have > to do a second purification step. > > -- > German Cancer Research Center (DKFZ) > Applied Tumorvirology - ATV F0200 > Christof Gaenzler > INF 242 > 69120 Heidelberg > T: +49-6221-42-4937 > F: +49-6221-42-4932 > > ----------------------------------------------------------------------- Petri Kursula "I am somehow less interested in the weight and University of Oulu convolutions of Einstein's brain than in the near Petri.Kursula@oulu.fi certainty that people of equal talent have lived http://start.at/MAG and died in cotton fields and sweatshops." http://cc.oulu.fi/~pkursula -- Stephen Jay Gould ----------------------------------------------------------------------- From owner-proteins@net.bio.net Tue Dec 14 16:06:02 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!outgoing.news.rcn.net.MISMATCH!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:06:51 +0000 Lines: 109 Message-ID: <385624CB.4DA3@erols.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: Sz62+7Gu7OTQ3jOihq6se/LppaHl7uHmh0A4zc7gtsE= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 14 Dec 1999 16:05:19 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:519 sci.med.nutrition:10255 misc.fitness.weights:51546 Siemel B. Naran wrote: > > On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse > > >Without all eight of the essential amino acids present, your body CANNOT do > >anything with that protein except break it down to synthesize the > >nonessential proteins and treat any surplus like fat. > > Well, the body may burn some muscle to make up for the difference (ie, for > whatever amino acids are lacking). Then the protein you eat is fully usable. > But of course, we don't want this. > > >Whey | Soy | Egg | Rice | EAA > >-----+-----|-----|------|---------- > > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > >If methionine is the limiting factor, you'll need over twice as much soy > >protein as you'd need whey protein to get the same usable EAA. > > > >Look at lysine, and you'll see that you need about three times as much rice > >as you'd need whey. > > If you eat twice as much soy, you get twice as much of every amino acid. > Which means too much protein, and therefore lots of work on your kidneys > to get rid of the excess protein/nitrogen. Thanks to the terrible job of health education that government run, tax supported, "public" schools are doing, you have been misinformed. The reason why they gave meat and dairy products their own food groups was to help livestock farmers. Until you buy a calorie counter that lists the amounts of amino acids in different foods, and until you go to Walton Feeds website, you just won't know what you're talking about. It's not the ratios of different amino acids in your blood that matter. You have enough of each amino acid in your blood to form the right proteins for tissue growth. Eight of these must be supplied by the food you eat, thirteen can be synthesized by your body if your food doesn't provide enough of them, and nutritionists currently classify histidine, the twenty-second amino acid, as only being essential for infants, meaning that babies need to get it from breastmilk or infant formula, but the rest of us don't have to worry about it. Now, in order to avoid muscle atrophy (the first noticible symptom of a protein deficiency) you need to replenish your amino acid pool on a regular basis. To do this, all you have to do is eat at leaast two meals a day, maintain your body weight, avoid eating abnormally large quantities of sugary fruit, and restrict your consumption of junk food. By the way, the chart I am talking about tells how many calories of any given food a 174 pound man (that's about average in the US) would have to eat to get his RDA of all eight amino acids. After looking at the chart, a person realizes that it is not neccessary for even the strictest vegetarian to carefully combine proteins. In fact, vegetarians and meat eaters alike are getting way too much protein. > >Some foods are heavier in sources than others. Since I want to rely largely > >on soy sources for protein for the time being, I'm trying to find vegan > >foods which are balanced in favor of lysine, methionine, threonine and > >tryptophan to make up the difference. According to the chart, if you weigh 174 pounds and you are an adult, you would only need to eat 304 calories of soybeans to get enough of each amino acid. If you eat fake meat made with soy protein, that number would be even lower because most of the carbohydrates and fat would be removed. I imagine you are smaller than 174 pounds, anyway. Peas and lentils are supposedly > >heavier in these, so the combination of peas and soy protein may result in > >something like whey's balance. No, you don't need to balance amino acids. Read the American Dietetic Association's position paper on vegetarianism. > > Grains are high in methionine, and maybe the others. So wheat cereal and > soy milk is probably a good choice. If you ate nothing but wheat and fit the description above, you would need 1061 calories to get your amino acids. I use tofu in my spaghetti (because > it tastes good to me), and this might be balance too because tofu is soy > and sphaghetti is grain. Combining proteins is like combining vitamins to get a "complete B Complex" or "complete tocopherol." > > >Mind you - that's -whey's- balance. I'm still looking for a source of > >information on what ratios a body really needs. Adults need 12mg of Isoleucine per kg of body weight, 16mg of Leucine per kg, 12mg of Lysine per kg, 10mg per kg of Methionine and/or Cystine, 16mg per kg of Phenylalanine and or Tyrosine, 8mg per kg of Threonine, 3mg per kg of body weight of Tryptophan, and 14mg per kg of Valine. One kg is about a pound. It may turn out that > >tryptophan or one of the others isn't needed in as great a proportion. > > I think whole egg has the balance the body needs. I think you're refering to protein effeciency ratios, and yes, eggs do have high protein effeciency ratios. Plant foods have low efficiency ratios when eaten alone, but when you combine them with the other foods eaten at the same meal it the ratio improves, and even if you only eat one plant food per meal you would still be fine as long as you didn't consistantly choose watermelon, apples, kiwis, etc. I looked hard on > the internet for a profile but couldn't find one (all I get are links > to products and info that I already know -- what the essential amino > acids are and what each one supposedly does). Anyway, as whey protein > has a BV of over 100 on rats, I'd say it has the right ratios. > > -- > -------------- > siemel b naran > -------------- From owner-proteins@net.bio.net Tue Dec 14 16:14:16 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!outgoing.news.rcn.net.MISMATCH!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:15:33 +0000 Lines: 118 Message-ID: <385626D5.4786@erols.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> <3855C83B.B018D5AD@internet.look.ca> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: d7MYpRRHszmpFpPVFVp0EA6v8YKrIB/DDLhW8+o3wlE= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 14 Dec 1999 16:13:58 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:520 sci.med.nutrition:10256 misc.fitness.weights:51550 Tom Matthews wrote: > > Am I the only one who has noticed that the level of discourse on this > newsgroup (smn) is at an all-time low? > > "Siemel B. Naran" wrote: > > > > On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse > > > > >Without all eight of the essential amino acids present, your body CANNOT do > > >anything with that protein except break it down to synthesize the > > >nonessential proteins and treat any surplus like fat. > > > > Well, the body may burn some muscle to make up for the difference (ie, for > > whatever amino acids are lacking). Then the protein you eat is fully usable. > > That is a really asinine answer! > That is like saying that you don't need to eat at all because you can > just digest your body from the inside! > > > But of course, we don't want this. > > Do you really need to say that? > > > >Whey | Soy | Egg | Rice | EAA > > >-----+-----|-----|------|---------- > > > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > > >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > > > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > > > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > > > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > > > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > > > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > > > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > > > >If methionine is the limiting factor, you'll need over twice as much soy > > >protein as you'd need whey protein to get the same usable EAA. > > > > > >Look at lysine, and you'll see that you need about three times as much rice > > >as you'd need whey. > > > > If you eat twice as much soy, you get twice as much of every amino acid. > > Which means too much protein, and therefore lots of work on your kidneys > > to get rid of the excess protein/nitrogen. > > "Lots of work" does not equal health problems or too much protein! > It has never been shown that high protein diets harm healthy kidneys. > Please learn some facts before answering! Actually, I have. Protein intakes far above 100g increase the risk of kidney failure, and certain types of protein, such as egg white protein and casein, increase the amount of calium that is removed from the bloodstream by the kidneys. In extreme cases, this causes kidney stones. If that calcium that is lost is not properly replaced (from milk, calcium pills, or food), osteoperosis will result. > > > >Some foods are heavier in sources than others. Since I want to rely largely > > >on soy sources for protein for the time being, > > It is unwise to rely on single or even restricted numbers of sources of > all vital nutrients. That's true. A healthy diet contains both starchy and nonstarchy vegetables, sweet raw fruits, grains, moderate amounts of bread and pasta, and a reliable source of active vitamin B12. > > > >I'm trying to find vegan > > >foods which are balanced in favor of lysine, methionine, threonine and > > >tryptophan to make up the difference. Peas and lentils are supposedly > > >heavier in these, so the combination of peas and soy protein may result in > > >something like whey's balance. > > No, the sulfur containing amino acids (methionine/cysteine) will still > be limiting. Last I heard, most people are actually getting too much sulpher. > Grains are what you need to balance beans/legumes. Read my other response. > > > Grains are high in methionine, and maybe the others. So wheat cereal and > > soy milk is probably a good choice. I use tofu in my spaghetti (because > > it tastes good to me), and this might be balance too because tofu is soy > > and sphaghetti is grain. > > Now you are talking. > But is it still nowhere as good an amino acid profile as > fish/meat/dairy/whole egg is. And it doesn't have to be. Current recommendations for protein intake are more than twice as high as most people need. > > > >Mind you - that's -whey's- balance. I'm still looking for a source of > > >information on what ratios a body really needs. It may turn out that > > >tryptophan or one of the others isn't needed in as great a proportion. > > Just fine a table of the RDAs for the essential amino acids and you will > have a rough indication of the ratio of body needs. > Sorry, I don't know where one is on-line and I don't have time to type > it in. > > > I think whole egg has the balance the body needs. I looked hard on > > the internet for a profile but couldn't find one (all I get are links > > to products and info that I already know -- what the essential amino > > acids are and what each one supposedly does). Anyway, as whey protein > > has a BV of over 100 on rats, I'd say it has the right ratios. > > Whole egg is one of the best available. > Actually (as I have said before) whole human is probably best, whole > monkey next and whole cow, whole chicken or whole fish not two bad. > Animal muscle meat only is not that great, because we make more than > simply muscle proteins. > But if you are body building animal muscle meat may be best of all. > > --Tom > Tom Matthews > > The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 > A non-profit membership organization dedicated to the extension > of the healthy human lifespan through ground breaking research, > innovative ideas and practical methods. > LIFE EXTENSION MAGAZINE - The ultimate source for new > health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 16:17:04 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.maxwell.syr.edu!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:17:49 +0000 Lines: 35 Message-ID: <3856275D.7A75@erols.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: 3zKsJSQwKkl+3Ku/VP7WDwpmVKdSOQoIyXZ/k1XHfoE= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 14 Dec 1999 16:16:14 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:521 sci.med.nutrition:10257 misc.fitness.weights:51551 Siemel B. Naran wrote: > > On Mon, 13 Dec 1999 10:33:23 -0600, River Mouse > > >Whey | Soy | Egg | Rice | EAA > >-----+-----|-----|------|---------- > > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > >10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > And L-Histidine is essential for children. > > But I'm thinking, does the body need an absolute amount of protein everyday > or an absolute ratio? > > If your body needs a fixed amount everyday, like 2.5 grams of methionine, > then you could just eats of lots of soy and you'd surely get or exceed the > minimum requirement of all the proteins. But through excess, you'd > overtax your kidneys in getting rid of the nitrogen. > > If your body needs a fixed ratio everyday, like lysine/methionine should > be 3.8 even though amount of methionine may be 1.5 or 2.5 or 3.5 grams, > then no amount of soy will satisfy the requirement. Recent studies show that soy protein does not increase calcium loss, but animal protein does. > > -- > -------------- > siemel b naran > -------------- From owner-proteins@net.bio.net Tue Dec 14 16:21:14 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!news.maxwell.syr.edu!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:22:25 +0000 Lines: 97 Message-ID: <38562870.365D@erols.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <833766$35e$1@flood.xnet.com> <3855C3DF.1BA33C6@internet.look.ca> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: AKQJSXdhueSD+g87XoTk0aPJSa3kGjXUnM2/cXaLYGg= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 14 Dec 1999 16:20:48 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:522 sci.med.nutrition:10258 misc.fitness.weights:51554 Tom Matthews wrote: > > You see, Stuart! > Even the person asking the question knows more than you! > Talk about the "blind leading the blind"!!! > > --Tom > > River Mouse wrote: > > > > "Stuart Dunn" wrote in message > > news:3854C135.2428@erols.com... > > > River Mouse wrote: > > > > > > > > I'm reading that without an adequate balance of the eight essential > > amino > > > > acids, the body is incapable of synthesizing tissue from protein. > > Instead, > > > > surplus protein is reduced to simpler proteins for synthesizing > > > > non-essential amino acids and/or converted to glucose and potentially to > > > > fat. > > [...] > > > > What ratio are the eight essential proteins needed in? Do these ratios > > > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > > > > > Where can I find reference for the levels of the essential amino acids > > > > contained in common foods? My particular interest is in vegan foods. > > > > > > It's not the ratios you need to be concerned with, it's the amounts. As > > > a rule of thumb, anyone eating at least 45 grams of protein a day from > > > sources other than gelatin and watermelon is safe, even if they have > > > active lifestyles. Except for sweets and greasy foods, all commonly > > > eaten vegan foods have "complete" proteins. Go to Walton Feed's website. > > > It has more details. > > > > This is a grossly inaccurate oversimplification. I know for a fact that many > > foods are lacking in some of the 8 essential amino acids You've been lied to. Meat producers were involved in designing nutrition education curriculums for public schools and they still advertise in publications geared toward nutritionists. I know, because I read those publications at Fenwick Library at George Mason University. If you look at a calorie counter you will see that all foods other than sweeteners, oils, and cornstarch have every amino acid. - these are the > > proteins which your body CANNOT synthesize on its own. > > > > Without all eight of the essential amino acids present, your body CANNOT do > > anything with that protein except break it down to synthesize the > > nonessential proteins and treat any surplus like fat. > > > > Let's say you take 100g of each of these four protein supplements (I'm > > listing supplements because I can't find Essential Amino Acid ratios for > > conventional foods yet). > > > > Whey: Cross-Flow Microfiltration Whey Protein Isolate > > Soy: 90% Soy Protein Isolate > > Egg: Egg White Protein Concentrate > > Rice: Rice Protein Concentrate > > > > Here's how many grams of each of these amino acids you'd get: > > > > (You might want to change to a fixed width font for this table) > > > > Whey | Soy | Egg | Rice | EAA > > -----+-----|-----|------|---------- > > 6.8 | 4.3 | 4.5 | 4.2 | L-Isoleucine > > 10.9 | 7.2 | 6.8 | 8.6 | L-Leucine > > 9.5 | 5.5 | 5.5 | 3.5 | L-Lysine > > 2.5 | 1.2 | 2.7 | 2.4 | L-Methionine > > 3.1 | 4.6 | ? | 5.2 | L-Phenylalanine > > 8.3 | 3.3 | 3.6 | 3.6 | L-Threonine > > 2.0 | 1.1 | 0.9 | 1.3 | L-Tryptophan > > 6.4 | 4.4 | 5.1 | 4.7 | L-Valine > > > > If methionine is the limiting factor, you'll need over twice as much soy > > protein as you'd need whey protein to get the same usable EAA. > > > > Look at lysine, and you'll see that you need about three times as much rice > > as you'd need whey. > > > > Some foods are heavier in sources than others. Since I want to rely largely > > on soy sources for protein for the time being, I'm trying to find vegan > > foods which are balanced in favor of lysine, methionine, threonine and > > tryptophan to make up the difference. Peas and lentils are supposedly > > heavier in these, so the combination of peas and soy protein may result in > > something like whey's balance. > > > > Mind you - that's -whey's- balance. I'm still looking for a source of > > information on what ratios a body really needs. It may turn out that > > tryptophan or one of the others isn't needed in as great a proportion. > > The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 > A non-profit membership organization dedicated to the extension > of the healthy human lifespan through ground breaking research, > innovative ideas and practical methods. > LIFE EXTENSION MAGAZINE - The ultimate source for new > health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 16:26:07 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!howland.erols.net!outgoing.news.rcn.net.MISMATCH!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:26:48 +0000 Lines: 55 Message-ID: <38562977.44A4@erols.com> References: <831b0q$u9$1@flood.xnet.com> <3854C135.2428@erols.com> <3855C34C.F5B6CF8D@internet.look.ca> Mime-Version: 1.0 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit X-Trace: aB1OVjYDv5m+j71YEO1wzgtwT0z4dgyIGv67pwZiKac= X-Complaints-To: abuse@rcn.com NNTP-Posting-Date: 14 Dec 1999 16:25:11 GMT X-Mailer: Mozilla 3.01-C-EROLS33599 (Macintosh; I; 68K) Xref: hgmp.mrc.ac.uk bionet.molbio.proteins:523 sci.med.nutrition:10259 misc.fitness.weights:51555 Tom Matthews wrote: > > Stuart Dunn wrote: > > > > River Mouse wrote: > > > > > > I'm reading that without an adequate balance of the eight essential amino > > > acids, the body is incapable of synthesizing tissue from protein. Instead, > > > surplus protein is reduced to simpler proteins for synthesizing > > > non-essential amino acids and/or converted to glucose and potentially to > > > fat. > > > > > > Another poster suggested that an excessive surplus could also result in the > > > formation of ill-defined muscle with the potential to break down more > > > easily. (Bulky versus lean muscle). > > > > > > What ratio are the eight essential proteins needed in? Do these ratios > > > change in repairing muscles under extreme duress, i.e. in weightlifting? > > > > > > Where can I find reference for the levels of the essential amino acids > > > contained in common foods? My particular interest is in vegan foods. > > It's not the ratios you need to be concerned with, it's the amounts. As > > a rule of thumb, anyone eating at least 45 grams of protein a day from > > sources other than gelatin and watermelon is safe, even if they have > > active lifestyles. Except for sweets and greasy foods, all commonly > > eaten vegan foods have "complete" proteins. Go to Walton Feed's website. > > It has more details. > > Stuart, since you obviously do not know what you are talking about, > would you please stop jumping in to answer questions with highly > incorrect answers! I heard the 45g figure about ten years ago when my parents were putting my little sister on a diet. They got a long appointment with a nutritionist who told them that 45g a day was adequate even if she got the protein from plant sources, even though she was still growing! Keep in mind that kids need far more protein than a normal adult who is the same size. Also, she said that protein complementation is unneccessary. If you go to the website I mentioned with the list of different foods, you will see that 45 grams is more than adults need. > > This is a science newsgroup, not a rumor mill! > > Please lurk for a while, read some texts, and solidify and correct your > knowledge for a while instead of leading innocent people astray. That's what I should be telling you. > > --Tom > Tom Matthews > > The LIFE EXTENSION FOUNDATION - http://www.lef.org - 800-544-4440 > A non-profit membership organization dedicated to the extension > of the healthy human lifespan through ground breaking research, > innovative ideas and practical methods. > LIFE EXTENSION MAGAZINE - The ultimate source for new > health and medical findings from around the world. From owner-proteins@net.bio.net Tue Dec 14 16:28:04 1999 Path: hgmp.mrc.ac.uk!server1.netnews.ja.net!fu-berlin.de!newsfeed.mathworks.com!cyclone.nyroc.rr.com!news.nyroc.rr.com!chnws02.mediaone.net!207.172.3.37!feed1.news.rcn.net!rcn!not-for-mail From: Stuart Dunn Newsgroups: alt.food.vegan,bionet.molbio.proteins,sci.med.nutrition,misc.fitness.weights Subject: Re: Essential amino acid requirements Date: Tue, 14 Dec 1999 11:29:12 +0000 Lines: 29 Message-ID: <38562A08.4A8E@erols.com> References: <831b0q$u9$1@flood.xnet.com> <01bf455f$9a1154e0$994065cb@default> <3854C21D.4A7D@erols.com> <81854.19750$eh2.347809@news2.rdc1.on.home.com> Mime-Version: 1.0 Content-Ty