Open AccessTarget Identification by Diazirine Photo‐Cross‐Linking and Click Chemistry
Author(s) -
MacKin Andrew L.,
Taunton Jack
Publication year - 2009
Publication title -
current protocols in chemical biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.503
H-Index - 14
ISSN - 2160-4762
DOI - 10.1002/9780470559277.ch090167
Subject(s) - diazirine , click chemistry , identification (biology) , chemistry , biophysics , combinatorial chemistry , biochemistry , botany , biology
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.