Unit

Target Identification by Diazirine Photo-Cross-Linking and Click Chemistry

  1. Andrew L. MacKinnon1,
  2. Jack Taunton1,2

Published Online: 1 DEC 2009

DOI: 10.1002/9780470559277.ch090167

Current Protocols in Chemical Biology

Current Protocols in Chemical Biology

How to Cite

MacKinnon, A. L. and Taunton, J. 2009. Target Identification by Diazirine Photo-Cross-Linking and Click Chemistry. Current Protocols in Chemical Biology. 1:55–73.

Author Information

  1. 1

    Program in Chemistry and Chemical Biology and Department of Cellular and Molecular Pharmacology, University of California San Francisco, San Francisco, California

  2. 2

    Howard Hughes Medical Institute, University of California San Francisco, San Francisco, California

Publication History

  1. Published Online: 1 DEC 2009

Abstract

Target identification is often the rate-determining step in deciphering the mechanism of action of biologically active small molecules. Photo-affinity labeling (PAL) represents a useful biochemical strategy for target identification in complex protein mixtures. This unit describes the use of alkyl diazirine-based photo-affinity probes and Cu(I)-catalyzed click chemistry to covalently label and visualize the targets of biologically active small molecules. A general method for affinity purification of probe-modified proteins, useful for identification of protein targets, is also described. Curr. Protoc. Chem Biol. 1:55-73. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • photo-affinity labeling;
  • diazirine;
  • click chemistry;
  • target identification;
  • affinity purification