Affinity proteomics to study endogenous protein complexes: Pointers, pitfalls, preferences and perspectives. (March 2015)

• Record Type: Journal Article
• Authors: LaCava, John; Molloy, Kelly R.; Taylor, Martin S.; Domanski, Michal; Chait, Brian T.; Rout, Michael P.
• Is Part Of: Biotechniques. Volume 58:Number 3(2015)
• Page Start: 103
• Page End: 119
• Publication Date: 2015-03
• Subjects: protein complex -- protein purification -- affinity -- proteomics -- interactomics; Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000114262
• Languages: English
• ISSNs: 0736-6205

Abstract

Dissecting and studying cellular systems requires the ability to specifically isolate distinct proteins along with the co-assembled constituents of their associated complexes. Affinity capture techniques leverage high affinity, high specificity reagents to target and capture proteins of interest along with specifically associated proteins from cell extracts. Affinity capture coupled to mass spectrometry (MS)-based proteomic analyses has enabled the isolation and characterization of a wide range of endogenous protein complexes. Here, we outline effective procedures for the affinity capture of protein complexes, highlighting best practices and common pitfalls.