Two Steps Forward--One Step Back: Advances in Affinity Purification Mass Spectrometry of Macromolecular Complexes

Proteomics. 2012 May;12(10):1591-608. doi: 10.1002/pmic.201100509.

Abstract

Cellular functions are defined by the dynamic interactions of proteins within macromolecular networks. Deciphering these complex interplays is the key to getting a comprehensive picture of cellular behavior and to understanding biological systems, from a simple bacterial cell to highly regulated neuronal cells or cancerous tissue. In the last decade, affinity purification (AP) coupled to mass spectrometry has emerged as a powerful tool to comprehensively study interaction networks and their macromolecular assemblies. This review discusses recent advances in AP approaches, from cell lysis to the importance of sample preparation and the choice of AP matrix as well as the development of different epitope tags and strategies to study dynamic interactions, with an emphasis on RNA-protein interaction networks.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Affinity Labels
  • Chromatography, Affinity / methods*
  • Mass Spectrometry / methods*
  • Protein Interaction Mapping / methods*
  • Proteins / chemistry*
  • Proteins / metabolism*
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • Systems Biology / methods*

Substances

  • Affinity Labels
  • Proteins
  • RNA-Binding Proteins
  • Ribonucleoproteins