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

Détails

ID Serval
serval:BIB_D9C0C14DF03C
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Modeling the isoelectric focusing of peptides in an OFFGEL multicompartment cell.
Périodique
Journal of Proteome Research
Auteur(s)
Lam H.T., Josserand J., Lion N., Girault H.H.
ISSN
1535-3893 (Print)
ISSN-L
1535-3893
Statut éditorial
Publié
Date de publication
2007
Peer-reviewed
Oui
Volume
6
Numéro
5
Pages
1666-1676
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
Résumé
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.
Mots-clé
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
Création de la notice
09/11/2014 23:46
Dernière modification de la notice
03/03/2018 21:53
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