Exploring Kinase Cosubstrate Promiscuity: Monitoring Kinase Activity through Dansylation
Exploring Kinase Cosubstrate Promiscuity: Monitoring Kinase Activity through Dansylation
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DOI：
10.1002/cbic.200800393
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发表时间：
2009-01-26
期刊：
影响因子：
3.2
通讯作者：
Pflum, Mary Kay H.
中科院分区：
文献类型：
--
作者：
Green, Keith D.;Pflum, Mary Kay H.
Kinases catalyze protein phosphorylation, which is a key event in cell signalling. Importantly, kinases are targeted by a number of drugs in clinical trials for cancer, rheumatoid arthritis, and immunosuppression.[1] Because kinases and protein phosphorylation play fundamental roles in disease, methods to monitor kinase activity and substrates are needed.We recently described a phosphoprotein-labelling reaction that couples kinases with an analogue of the adenosine 5′-tri-phosphate (ATP, 1a) cosubstrate. Specifically, ATP conjugated to biotin through the γ-phosphate (ATP-biotin) served as a kinase cosubstrate, which allowed phosphoprotein biotinylation in vitro and in cell lysates.[2] The studies with ATP-biotin raise the possibility that cellular kinases generally tolerate γ-phosphate-modified ATP analogues (Scheme 1A) as cosubstrates. Cosubstrate promiscuity is documented with multiple protein-modifying enzymes, including farnesyltransferase,[3] transglutaminase,[4] galactosyltransferase,[5] N-acetyltransferase,[6] DNA methyltransferase,[7] and phosphopantetheinyl transferase (PPtase).[8] To explore kinase cosubstrate promiscuity, we hypothesized that additional γ-phosphate-modified ATP analogues would be compatible with kinase-catalyzed labelling.
