Purifying protein complexes for mass spectrometry: applications to protein translation

Methods. 2005 Mar;35(3):274-90. doi: 10.1016/j.ymeth.2004.08.019. Epub 2005 Jan 12.

Abstract

Proteins control and mediate most of the biological activities in the cell. In most cases, proteins either interact with regulatory proteins or function in large molecular assemblies to carryout biological processes. Understanding the functions of individual proteins requires the identification of these interacting proteins. With its speed and sensitivity, mass spectrometry has become the dominant method for identifying components of protein complexes. This article reviews and discusses various approaches to purify protein complexes and analyze the proteins using mass spectrometry. As examples, methods to isolate and analyze protein complexes responsible for the translation of messenger RNAs into polypeptides are described.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Differentiation
  • Centrifugation, Density Gradient / methods
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / chemistry
  • Hydrogen-Ion Concentration
  • Immunoprecipitation
  • Mass Spectrometry / methods*
  • Models, Biological
  • Protein Biosynthesis*
  • Proteins / chemistry*
  • Proteins / isolation & purification*
  • Proteomics / methods*
  • RNA, Messenger / metabolism
  • Ribosomes / chemistry
  • Saccharomyces cerevisiae / metabolism
  • Sensitivity and Specificity
  • Sucrose / pharmacology
  • Temperature
  • Time Factors

Substances

  • Epitopes
  • Proteins
  • RNA, Messenger
  • Sucrose