Modeling the isoelectric focusing of peptides in an OFFGEL multicompartment cell.

Details

Serval ID
serval:BIB_D9C0C14DF03C
Type
Article: article from journal or magazin.
Collection
Publications
Title
Modeling the isoelectric focusing of peptides in an OFFGEL multicompartment cell.
Journal
Journal of Proteome Research
Author(s)
Lam H.T., Josserand J., Lion N., Girault H.H.
ISSN
1535-3893 (Print)
ISSN-L
1535-3893
Publication state
Published
Issued date
2007
Peer-reviewed
Oui
Volume
6
Number
5
Pages
1666-1676
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Abstract
In proteomic analysis of complex samples at the peptide level (termed shotgun proteomics), an effective prefractionation is crucial to decrease the complexity of the peptide mixture for further analysis. In this perspective, the high-resolving power of the IEF fractionation step is a determining parameter, in order to obtain well-defined fractions and correct information on peptide isoelectric points, to provide an additional filter for protein identification. Here, we explore the resolving power of OFFGEL IEF as a prefractionation tool to separate peptides. By modeling the peak width evolution versus the peptide charge gradient at pI, we demonstrate that for the three proteomes considered in silico (Deinococcus radiodurans, Saccharomyces cerevisiae, and Homo sapiens), 90% of the peptides should be correctly focused and recovered in two wells at most. This result strongly suggests OFFGEL to be used as a powerful fractionation tool in shotgun proteomics. The influence of the height and shape of the compartments is also investigated, to give the optimal cell dimensions for an enhanced peptide recovery and fast focusing time.
Keywords
Deinococcus/chemistry, Humans, Isoelectric Focusing/instrumentation, Isoelectric Focusing/methods, Models, Theoretical, Peptides/chemistry, Peptides/genetics, Proteome/analysis, Reproducibility of Results, Saccharomyces cerevisiae/chemistry
Pubmed
Web of science
Create date
09/11/2014 22:46
Last modification date
20/08/2019 15:59
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