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• W1996817672 abstract "We developed a new approach to distinguish distinct protein conformations in live cells. The method, exposable tetracysteine (XTC), involved placing an engineered tetracysteine motif into a target protein that has conditional access to biarsenical dye binding by conformational state. XTC was used to distinguish open and closed regulatory conformations of Src family kinases. Substituting just four residues with cysteines in the conserved SH2 domain of three Src-family kinases (c-Src, Lck, Lyn) enabled open and closed conformations to be monitored on the basis of binding differences to biarsenical dyes FlAsH or ReAsH. Fusion of the kinases with a fluorescent protein tracked the kinase presence, and the XTC approach enabled simultaneous assessment of regulatory state. The c-Src XTC biosensor was applied in a boutique screen of kinase inhibitors, which revealed six compounds to induce conformational closure. The XTC approach demonstrates new potential for assays targeting conformational changes in key proteins in disease and biology." @default.
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• W1996817672 date "2014-05-15" @default.
• W1996817672 modified "2023-10-18" @default.
• W1996817672 title "A Biosensor of Src Family Kinase Conformation by Exposable Tetracysteine Useful for Cell-Based Screening" @default.
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• W1996817672 doi "https://doi.org/10.1021/cb500242q" @default.
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