Tandem Affinity Purification Combined with Mass Spectrometry to Identify Components of Protein Complexes
Open Access
- 1 January 2008
- book chapter
- research article
- Published by Springer Science and Business Media LLC
- Vol. 439, 309-326
- https://doi.org/10.1007/978-1-59745-188-8_21
Abstract
Most biological processes are governed by multiprotein complexes rather than individual proteins. Identification of protein complexes therefore is becoming increasingly important to gain a molecular understanding of cells and organisms. Mass spectrometry—based proteomics combined with affinity-tag-based protein purification is one of the most effective strategies to isolate and identify protein complexes. The development of tandem-affinity purification approaches has revolutionized proteomics experiments. These two-step affinity purification strategies allow rapid, effective purification of protein complexes and, at the same time, minimize background. Identification of even very low-abundant protein complexes with modern sensitive mass spectrometers has become routine. Here, we describe two general strategies for tandem-affinity purification followed by mass spectrometric identification of protein complexes.Keywords
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