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. 2012:917:391-407.
doi: 10.1007/978-1-61779-992-1_22.

Complementary proteomic analysis of protein complexes

Todd M Greco  1 , Yana Miteva, Frank L Conlon, Ileana M Cristea
Affiliations

Complementary proteomic analysis of protein complexes

Todd M Greco et al. Methods Mol Biol. 2012.

Abstract

Proteomic characterization of protein complexes leverages the versatile platform of liquid chromatography-tandem mass spectrometry to elucidate molecular and cellular signaling processes underlying the dynamic regulation of macromolecular assemblies. Here, we describe a complementary proteomic approach optimized for immunoisolated protein complexes. As the relative complexity, abundance, and physiochemical properties of proteins can vary significantly between samples, we have provided (1) complementary sample preparation workflows, (2) detailed steps for HPLC and mass spectrometric method development, and (3) a bioinformatic workflow that provides confident peptide/protein identification paired with unbiased functional gene ontology analysis. This protocol can also be extended for characterization of larger complexity samples from whole cell or tissue Xenopus proteomes.

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Figures

Fig. 1
Fig. 1
Workflow summary for proteomic analysis of immunoisolated protein complexes; diagram of LTQ-Orbitrap Velos from Thermo Fisher Scientific.
See this image and copyright information in PMC

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