Complementary proteomic analysis of protein complexes
Publication Year
2012
Type
Journal Article
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.
Keywords
Animals,
proteomics,
Molecular Sequence Data,
Amino Acid Sequence,
Multiprotein Complexes,
Proteome,
Xenopus Proteins,
Chromatography, High Pressure Liquid,
Tandem Mass Spectrometry,
Peptide Fragments,
Xenopus,
Electrophoresis, Polyacrylamide Gel,
Proteolysis,
Trypsin,
Peptide Mapping,
Reference Standards
Journal
Methods Mol Biol
Volume
917
Pages
391-407
ISSN Number
1940-6029
Alternate Journal
Methods Mol Biol
PMID
22956100