Standard Article

Limits of Proteomics: Protein Solubilisation Issues

  1. Thierry Rabilloud

Published Online: 15 MAR 2012

DOI: 10.1002/9780470015902.a0006201.pub2

eLS

eLS

How to Cite

Rabilloud, T. 2012. Limits of Proteomics: Protein Solubilisation Issues. eLS. .

Author Information

  1. CEA-Grenoble, iRTSV/LCBM, UMR CNRS-CEA-UJF 5249, Grenoble, France

Publication History

  1. Published Online: 15 MAR 2012

Abstract

Because of their tremendous chemical heterogeneity, many proteins, such as membrane proteins, are poorly soluble in aqueous solvents. This precludes their analysis by classical proteomics techniques, whether these techniques are based on protein separation (e.g. two-dimensional gel-based proteomics) or on peptide separation after in-solution digestion. For techniques based on peptide separation, the issue is to couple on the one side a complete solubilisation of proteins, a good efficiency of the protease used to produce the peptides, and on the other side minimal interference with the techniques used to separate the peptides. For techniques based on protein separation, the issue is to solubilise the proteins and to keep them soluble under the conditions used for the protein separation. For both type of technologies, combinations of detergents and chaotropes often provide the most performing solution to the protein solubilisation problem, although complete solubilisation can never be guaranteed.

Key Concepts:

  • Proteins must be extracted from their cellular context into an aqueous-based solvent for their analysis.

  • Surfactants/detergents provide the adequate medium to mimic a lipid-like environment in water.

  • The structural variety of detergents provides the various solubilisation performances that can be adapted to many proteomics methods.

  • Chaotropes are often used in conjunction with detergents to improve protein/peptide solubility.

Keywords:

  • proteomics;
  • detergents;
  • membrane;
  • chaotropes;
  • solubilisation