Digestion and depletion of abundant proteins improves proteomic coverage

Nat Methods. 2013 Jan;10(1):54-6. doi: 10.1038/nmeth.2250. Epub 2012 Nov 18.

Abstract

Two major challenges in proteomics are the large number of proteins and their broad dynamic range in the cell. We exploited the abundance-dependent Michaelis-Menten kinetics of trypsin digestion to selectively digest and deplete abundant proteins with a method we call DigDeAPr. We validated the depletion mechanism with known yeast protein abundances, and we observed greater than threefold improvement in low-abundance human-protein identification and quantitation metrics. This methodology should be broadly applicable to many organisms, proteases and proteomic pipelines.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Humans
  • Mass Spectrometry / methods*
  • Peptide Fragments / analysis*
  • Proteins / metabolism*
  • Proteome / analysis*
  • Proteomics / methods*

Substances

  • Peptide Fragments
  • Proteins
  • Proteome