Assays in which the detection of a biological phenomenon is coupled to the production of bioluminescence by luciferase have gained widespread use. As fi refl y luciferases (FLuc) and kinases share a common substrate (ATP), coupling of a kinase to FLuc allows for the amount of ATP remaining following a kinase reaction to be assessed by quantitating the amount of luminescence produced. Alternatively, the amount of ADP produced by the kinase reaction can be coupled to FLuc through a two-step process. This chapter describes the bioluminescent assays that were developed for three classes of kinases (lipid, protein, and metabolic kinases) and miniaturized to 1536-well format, enabling their use for quantitative highthroughput (qHTS) of small-molecule libraries.
CITATION STYLE
Davis, M. I., Auld, D. S., & Inglese, J. (2016). Bioluminescence methods for assaying kinases in quantitative high-throughput screening (qHTS) format applied to yes1 tyrosine kinase, glucokinase, and PI5P4Kα lipid kinase. Methods in Molecular Biology, 1360, 47–58. https://doi.org/10.1007/978-1-4939-3073-9_4
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