A simple and effective cleavable linker for chemical proteomics applications

Mol Cell Proteomics. 2013 Jan;12(1):237-44. doi: 10.1074/mcp.M112.021014. Epub 2012 Oct 1.

Abstract

The study of metabolically labeled or probe-modified proteins is an important area in chemical proteomics. Isolation and purification of the protein targets is a necessary step before MS identification. The biotin-streptavidin system is widely used in this process, but the harsh denaturing conditions also release natively biotinylated proteins and non-selectively bound proteins. A cleavable linker strategy is a promising approach for solving this problem. Though several cleavable linkers have been developed and tested, an efficient, easily synthesized, and inexpensive cleavable linker is a desirable addition to the proteomics toolbox. Here, we describe the chemical proteomics application of a vicinal diol cleavable linker. Through easy-to-handle chemistry we incorporate this linker into an activity-based probe and a biotin alkyne tag amenable for bioorthogonal ligation. With these reagents, background protein identifications are significantly reduced relative to standard on-bead digestion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / chemistry
  • Cathepsins
  • Cross-Linking Reagents / chemistry*
  • Glucosides / chemistry
  • Mass Spectrometry
  • Proteins
  • Proteome / analysis
  • Proteome / chemistry
  • Proteomics / methods*
  • Pyrimidinones / chemistry

Substances

  • Cross-Linking Reagents
  • Glucosides
  • Proteins
  • Proteome
  • Pyrimidinones
  • Biotin
  • vicine
  • Cathepsins