Complementary proteomic analysis of protein complexes

Todd M. Greco, Yana Miteva, Frank L. Conlon, Ileana M. Cristea

Research output: Chapter in Book/Report/Conference proceedingChapter

16 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationXenopus Protocols
Subtitle of host publicationPost-Genomic Approaches
PublisherHumana Press Inc.
Pages391-407
Number of pages17
ISBN (Print)9781617799914
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume917
ISSN (Print)1064-3745

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics

Keywords

  • Bioinformatics
  • HPLC
  • In-gel digestion
  • In-solution digestion
  • Mass spectrometry
  • Protein complex
  • Protein interactions
  • Proteomics
  • Xenopus

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