SEARCH

SEARCH BY CITATION

Keywords:

  • ATP;
  • cdk2;
  • chemical genetics;
  • cyclin;
  • kinases;
  • substrate isolation;
  • thiophosphorylation

Abstract

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 N6-(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.