A p38alpha-Selective Chemosensor for use in Unfractionated Cell Lysates
Author(s)Stains, Cliff I.; Lukovic, Elvedin; Imperiali, Barbara
A p38α-Selective Chemosensor for use in Unfractionated Cell Lysates
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Recent efforts have identified the p38alpha Ser/Thr kinase as a potential target for the treatment of inflammatory diseases as well as non-small cell lung carcinoma. Despite the significance of p38alpha, no direct activity probe compatible with cell lysate analysis exists. Instead, proxies for kinase activation, such as phosphospecific antibodies, which do not distinguish between p38 isoforms, are often used. Our laboratory has recently developed a sulfonamido-oxine (Sox) fluorophore that undergoes a significant increase in fluorescence in response to phosphorylation at a proximal residue, allowing for real-time activity measurements. Herein we report the rational design of a p38alpha-selective chemosensor using this approach. We have validated the selectivity of this sensor using specific inhibitors and immunodepletions and show that p38α activity can be monitored in crude lysates from a variety of cell lines, allowing for the potential use of this sensor in both clinical and basic science research applications.
DepartmentMassachusetts Institute of Technology. Department of Biology; Massachusetts Institute of Technology. Department of Chemistry
ACS Chemical Biology
American Chemical Society
Stains, Cliff I., Elvedin Luković, and Barbara Imperiali. “A p38alpha-Selective Chemosensor for Use in Unfractionated Cell Lysates.” ACS Chemical Biology 6.1 (2011): 101–105.
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