Method for qualitative comparisons of protein mixtures based on enzyme-catalyzed stable-isotope incorporation

Item Type:	Article
Title:	Method for qualitative comparisons of protein mixtures based on enzyme-catalyzed stable-isotope incorporation
Creators Name:	Mirgorodskaya, E. and Wanker, E.E. and Otto, A. and Lehrach, H. and Gobom, J.
Abstract:	Determining which proteins are unique among one or several protein populations is an often-encountered task in proteomics. To this purpose, we present a new method based on trypsin-catalyzed incorporation of the stabile isotope 18O in the C-termini of tryptic peptides, followed by LC-MALDI MS analysis. The analytical strategy was designed such that proteins unique to a given population out of several can be assigned in a single experiment by the isotopic signal intensity distributions of their tryptic peptides in the recorded mass spectra. The method is demonstrated for protein-protein interaction analysis, in which the differential isotope labeling was used to distinguish endogenous human brain proteins interacting with a recombinant bait protein from nonbiospecific background binders.
Keywords:	18 o Labeling, Comparative Proteomics, MALDI, Mass Spectrometry, Nano-Liquid Chromatography, Protein Interactions, Stable Isotope
Source:	Journal of Proteome Research
ISSN:	1535-3893
Publisher:	American Chemical Society
Volume:	4
Number:	6
Page Range:	2109-2116
Date:	1 November 2005
Official Publication:	https://doi.org/10.1021/pr050219i
PubMed:	View item in PubMed

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