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Complementary Proteomic Analysis of Protein Complexes

Author(s): Greco, Todd M.; Miteva, Yana; Conlon, Frank L; Cristea, Ileana M.

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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.
Publication Date: 1-Jan-2012
DOI: doi:10.1007/978-1-61779-992-1_22
EISSN: 1940-6029
Keywords: Xenopus, Proteomics, Protein complex, Protein interactions, In-gel digestion, In-solution digestion, Mass spectrometry, HPLC, Bioinformatics
Language: eng
Type of Material: Journal Article
Journal/Proceeding Title: Methods in Molecular Biology
Version: Author's manuscript

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