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

Authors

  • E. Mirgorodskaya
  • E.E. Wanker
  • A. Otto
  • H. Lehrach
  • J. Gobom

Journal

  • Journal of Proteome Research

Citation

  • J Proteome Res 4 (6): 2109-2116

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.


DOI

doi:10.1021/pr050219i