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The elucidation of signalling pathways relies heavily upon the identification of protein kinase substrates. Recent investigations have demonstrated the efficacy of chemical genetics using ATP analogues and modified protein kinases for specific substrate labelling. Here we combine N(6) -(cyclohexyl)ATPγS with an analogue-sensitive cdk2 variant to thiophosphorylate its substrates and demonstrate a pH-dependent, chemoselective, one-step alkylation to facilitate the detection or isolation of thiophosphorylated peptides.

Original publication

DOI

10.1002/cbic.201000528

Type

Journal article

Journal

Chembiochem

Publication Date

07/03/2011

Volume

12

Pages

633 - 640

Keywords

Alkylation, Chlorides, Cyclin-Dependent Kinase 2, Cysteine, Electrophoresis, Polyacrylamide Gel, Hydrogen-Ion Concentration, Peptides, Phosphorus Compounds, Signal Transduction, Substrate Specificity