Analyzing protein-protein interactions by quantitative mass spectrometry

Item Type:	Review
Title:	Analyzing protein-protein interactions by quantitative mass spectrometry
Creators Name:	Paul, F.E. and Hosp, F. and Selbach, M.
Abstract:	Since most cellular processes depend on interactions between proteins, information about protein-protein interactions (PPIs) provide valuable insights into protein function. Over the last years, quantitative affinity purification followed by mass spectrometry (q-AP-MS) has become a powerful approach to investigate PPIs in an unbiased manner. In q-AP-MS the protein of interest is biochemically enriched together with its interaction partners. In parallel, a control experiment is performed to control for non-specific binding. Quantitative mass spectrometry is then employed to compare protein levels in both samples and to exclude non-specific contaminants. Here, we provide two detailed q-AP-MS protocols for pull-downs with immobilized bait proteins or transient transfection of tagged expression constructs. We discuss benefits and limitations of q-AP-MS and highlight critical parameters that need to be considered. The protocols and background information presented here allow the reader to adapt the generic q-AP-MS strategy for a wide range of biological questions.
Keywords:	Protein-Protein Interactions, SILAC, Mass Spectrometry, Pull-Down, Immunoprecipitation
Source:	Methods
ISSN:	1046-2023
Publisher:	Elsevier / Academic Press
Volume:	54
Number:	4
Page Range:	387-395
Date:	August 2011
Official Publication:	https://doi.org/10.1016/j.ymeth.2011.03.001
PubMed:	View item in PubMed

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