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The chemoselective one-step alkylation and isolation of thiophosphorylated Cdk2 substrates in the presence of native cysteine
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Lee, Sarah E., Elphick, Lucy M., Kramer, Holger B., Jones, Alexandra M., Child, Emma S., Anderson, Alexandra A., Bonnac, Laurent, Suwaki, Natsuko, Kessler, Benedikt M., Gouverneur, Véronique and Mann, David J. (2011) The chemoselective one-step alkylation and isolation of thiophosphorylated Cdk2 substrates in the presence of native cysteine. ChemBioChem, Volume 12 (Number 4). pp. 633-640. doi:10.1002/cbic.201000528 ISSN 1439-4227.
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Official URL: http://dx.doi.org/10.1002/cbic.201000528
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.
Item Type: | Journal Article | ||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||
Journal or Publication Title: | ChemBioChem | ||||
Publisher: | Wiley - V C H Verlag GmbH & Co. KGaA | ||||
ISSN: | 1439-4227 | ||||
Official Date: | 7 March 2011 | ||||
Dates: |
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Volume: | Volume 12 | ||||
Number: | Number 4 | ||||
Page Range: | pp. 633-640 | ||||
DOI: | 10.1002/cbic.201000528 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published |
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