These authors contributed equally to this work.
The Chemoselective One-Step Alkylation and Isolation of Thiophosphorylated Cdk2 Substrates in the Presence of Native Cysteine
Article first published online: 8 FEB 2011
Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Volume 12, Issue 4, pages 633–640, March 7, 2011
How to Cite
Lee, S. E., Elphick, L. M., Kramer, H. B., Jones, A. M. E., Child, E. S., Anderson, A. A., Bonnac, L., Suwaki, N., Kessler, B. M., Gouverneur, V. and Mann, D. J. (2011), The Chemoselective One-Step Alkylation and Isolation of Thiophosphorylated Cdk2 Substrates in the Presence of Native Cysteine. ChemBioChem, 12: 633–640. doi: 10.1002/cbic.201000528
- Issue published online: 25 FEB 2011
- Article first published online: 8 FEB 2011
- Manuscript Received: 3 SEP 2010
- Cancer Research UK
- chemical genetics;
- substrate isolation;
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.