title: Target Identification by Diazirine Photo-Cross-linking and Click Chemistry.
abstract: Target identification of biologically active small-molecules is often the rate-determining step in forward chemical genetics. 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.
authors.affiliation: Program in Chemistry and Chemical Biology and Department of Cellular and Molecular Pharmacology University of California, San Francisco San Francisco, California 94158-2280, USA.
authors.forename: Andrew L
authors.initials: AL
authors.lastname: Mackinnon
authors.forename: Jack
authors.initials: J
authors.lastname: Taunton
keywords: affinity purification; click chemistry; diazirine; photo-affinity labeling; target identification