Complementary proteomic analysis of protein complexes.

Complementary proteomic analysis of protein complexes.
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DOI:
10.1007/978-1-61779-992-1_22
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发表时间:
2012-01-01
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
--
通讯作者:
Cristea, Ileana M
Cristea, Ileana M
中科院分区:
其他
文献类型:
--
作者:
Greco, Todd M;Miteva, Yana;Cristea, Ileana M

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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.