Probing proteomes with benzophenone photoprobes

Methods Mol Biol. 2012:803:65-75. doi: 10.1007/978-1-61779-364-6_6.

Abstract

Benzophenone photoprobes represent powerful tools for chemical proteomics. Upon UV irradiation, a benzophenone photoprobe can selectively form a covalent bond with its target protein in complex protein mixtures. Thus, photoprobes can be used to profile a wide variety of proteins in complex proteomes. This chapter describes simple protocols to derivatize fluorenylmethyloxycarbonyl (Fmoc)-protected peptide-nucleic-acid adenine (PNA adenine) into a benzophenone photoprobe and its application in photolabeling its target proteins. The method as described does not require specialized equipment for probe synthesis and photolabeling. In addition, the strategy is applicable to recognition motifs other than PNA adenine, such as peptides, to profile their target proteins in complex proteomes.

Publication types

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

MeSH terms

  • Adenine / metabolism
  • Benzophenones / metabolism*
  • Blotting, Western
  • Cell Extracts
  • Cytosol / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Jurkat Cells
  • Light*
  • Molecular Probes / chemistry
  • Molecular Probes / metabolism
  • Peptide Nucleic Acids / metabolism
  • Proteome / metabolism*
  • Proteomics / methods*
  • Solid-Phase Synthesis Techniques

Substances

  • Benzophenones
  • Cell Extracts
  • Molecular Probes
  • Peptide Nucleic Acids
  • Proteome
  • benzophenone
  • Adenine