AnGenMap

Sample Discussion

Subject: Porcine protein database search and GO annotations

From enjoy_your_dream1210_at_yahoo.co.jp  Tue Jul 12 09:04:56 2005
Date: Tue, 12 Jul 2005 21:19:14 +0900 (JST)
From: =?ISO-2022-JP?B?GyRCMGY+ZRsoQiAbJEJDaU06GyhC?= <enjoy_your_dream1210_at_yahoo.co.jp>
Subject: Porcine protein database
To: Multiple Recipients of <angenmap_at_animalgenome.org>

Dear Sri/Madam;

I am performing the proteomics research on porcine
development and currently have a problem with protein
identification after mass-spectrometry analysis of
trypsin-digested peptides.

I would like to know which database is the most suitable
for protein search (database maching)?
If anyone have an idea or advice, please give me any
information.
Thank you.

Very sincerely yours,
Tadao

Tadao Inoue, M. D.
985 Sanzo, Higashimura-cho,
Fukuyama, Hiroshima, 729-0292
Japan
TEL: +81-84-936-2112 (4628)
FAX: +81-84-936-2459
E-mail: enjoy_your_dream1210_at_yahoo.co.jp
From vallet_at_email.marc.usda.gov  Fri Jul 15 14:48:33 2005
From: "Jeff Vallet" <vallet_at_email.marc.usda.gov>
Subject: porcine protein database
Date: Fri, 15 Jul 2005 13:45:12 -0500
To: Multiple Recipients of <angenmap_at_animalgenome.org>

Dr. Inoue,

        When we do protein searches, we use the option that searches
against all mammalian sequences. We have tried using the porcine EST
sequences, but in our hands this information is not complete enough to
provide a useful search. Using the mammalian sequences option and Mascot
searching, for example, it is usually possible to get appropriate IDs.
However, mass based searching such as Mascot, will often give the
incorrect ID simply because the actual sequence of the protein is not
present in the database, because the swine sequence database is
incomplete, and homologous sequences from other species are not
sufficiently identical to achieve a high enough score to produce a
match. To deal with this problem, we do two types of searches, a Mascot
or mass based search, and a homology tolerant search using MS-homology
from protein prospector, which is available on the web. To do this type
of search, we use the PEAKS program from Bioinformatics Solutions, Inc.
to obtain de novo sequence information from the doubly charged peptides
from our tandem mass spectrometry analysis of a given spot. Then, we
submit all the de novo sequences from a particular spot to MS-homology
in a batch. The homology tolerant search is not as dependent on an exact
match as Mascot, and so it will often give an ID that is different from
that of Mascot. Two thirds of the time, Mascot and MS-homology give the
same ID, supporting the ID. For the other third of the time, we feel
that MS-homology search, which is more tolerant of mismatches, is
probably the better ID, but at least now have two alternative candidates
for further research. Further details of our use of homology tolerant
search methods can be found in Kayser et al., 2004, Journal of
Biomolecular Techniques, 15:285-295.

Jeffrey L. Vallet
Research Leader
USDA, ARS, Roman L. Hruska US Meat Animal Research Center
PO Box 166
Clay Center, NE 68933
402-762-4187
FAX 402-762-4382
From enjoy_your_dream1210_at_yahoo.co.jp  Thu Feb 16 01:23:19 2006
Date: Thu, 16 Feb 2006 16:23:19 +0900 (JST)
From: =?ISO-2022-JP?B?GyRCMGY+ZRsoQiAbJEJDaU06GyhC?= <enjoy_your_dream1210_at_yahoo.co.jp>
Subject: Porcine protein annotation
To: Multiple Recipients of <angenmap_at_animalgenome.org>

Hi, everyone, 

I am currently working on porcine proteome and handling hundreds of proteins
of interest. I would like to categorize them into several groups on
functional annotation (for example, protein synthesis group, metabolism
group, membrane transport group, etc). Could anyone kindly please show me
how this could be done?

Very sincerely yours, Tadao

==========================================
Tadao Inoue 
Department of Biotechnology, 
Fukuyama University 
985 Sanzo, Higashimura-cho, 
Fukuyama, Hiroshima, 729-0292 
Japan 
TEL: +81-84-936-2112 (4628) 
FAX: +81-84-936-2459 
E-mail: enjoy_your_dream1210_at_yahoo.co.jp 
=========================================== 
From zhu_at_iastate.edu  Thu Feb 16 08:27:56 2006
Date: Thu, 16 Feb 2006 08:27:51 -0600
From: Zhiliang Hu <zhu_at_iastate.edu>
Subject: Re: Porcine protein annotation
To: Multiple Recipients of <angenmap_at_animalgenome.org>

Tadao, 

If you can blast your proteins against the GO peptide db to obtain the GO
ids for them, you can use the GO database/tools (http://www.geneontology.org/;
"AmiGO", for example) to categorize them into "functional annotation" of your
choice. If what you wanted was to get a simple count of appearance of your
proteins within each of the "functional annotation", you can use my recently
developed (http://www.animalgenome.org/bioinfo/resources/util/countgo.html)
on-line tool -- currently this tool supports several commonly used "GO Slim"
classifications such as GO_Slim, GOA, EGAD, etc.

Good luck,

Zhiliang
/NAGRP Bioinformatics
 
At 01:23 AM 2/16/2006, =?ISO-2022-JP?B?GyRCMGY+ZRsoQiAbJEJDaU06GyhC?= wrote: 

> Hi, everyone,
>
> I am currently working on porcine proteome and handling hundreds of proteins
> of interest. I would like to categorize them into several groups on
> functional annotation (for example, protein synthesis group, metabolism
> group, membrane transport group, etc). Could anyone kindly please show me
> how this could be done?
>
> Very sincerely yours,
>
> Tadao

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