Abstract
Assays in which the detection of a biological phenomenon is coupled to the production of bioluminescence by luciferase have gained widespread use. As firefly 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 high-throughput (qHTS) of small-molecule libraries.
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References
Cohen P, Alessi DR (2012) Kinase drug discovery—what’s next in the field? ACS Chem Biol 8:96–104
Walsh MJ, Brimacombe KR, Veith H et al (2011) 2-Oxo-N-aryl-1,2,3,4-tetrahydroquinoline-6-sulfonamides as activators of the tumor cell specific M2 isoform of pyruvate kinase. Bioorg Med Chem Lett 21:6322–6327
Harbert C, Marshall J, Soh S, Steger K (2008) Development of a HTRF kinase assay for determination of Syk activity. Curr Chem Genomics 1:20–26
Fabian MA, Biggs WH 3rd, Treiber DK et al (2005) A small molecule-kinase interaction map for clinical kinase inhibitors. Nat Biotechnol 23:329–336
Kleman-Leyer KM, Klink TA, Kopp AL et al (2009) Characterization and optimization of a red-shifted fluorescence polarization ADP detection assay. Assay Drug Dev Technol 7:56–67
Hastie CJ, McLauchlan HJ, Cohen P (2006) Assay of protein kinases using radiolabeled ATP: a protocol. Nat Protoc 1:968–971
Tanega C, Shen M, Mott BT, Thomas CJ, MacArthur R, Inglese J, Auld DS (2009) Comparison of bioluminescent kinase assays using substrate depletion and product formation. Assay Drug Dev Technol 7:606–614
Somberg R, Goueli SA (2004) Method for detecting transferase enzymatic activity. In US 2004/0101922 A1 (USPTO ed., Promega Corporation, USA
Wu G, Yuan Y, Hodge CN (2003) Determining appropriate substrate conversion for enzymatic assays in high-throughput screening. J Biomol Screen 8:694–700
Segel IH (1993) Enzyme kinetics: behavior and analysis of rapid equilibrium and steady-state enzyme systems. John Wiley and Sons, New York
Li H, Totoritis RD, Lor LA, Schwartz B, Caprioli P, Jurewicz AJ, Zhang G (2009) Evaluation of an antibody-free ADP detection assay: ADP-Glo. Assay Drug Dev Technol 7:598–605
Sanghera J, Li R, Yan J (2009) Comparison of the luminescent ADP-Glo assay to a standard radiometric assay for measurement of protein kinase activity. Assay Drug Dev Technol 7:615–622
Zegzouti H, Goueli SA (2010) ADP Detection based luminescent phosphotransferase or ATP hydrolase assay. In US 2010/0075350 A1 (USPTO ed., Promega Corporation, USA
Auld DS, Southall NT, Jadhav A, Johnson RL, Diller DJ, Simeonov A, Austin CP, Inglese J (2008) Characterization of chemical libraries for luciferase inhibitory activity. J Med Chem 51:2372–2386
Auld DS, Zhang YQ, Southall NT, Rai G, Landsman M, MacLure J, Langevin D, Thomas CJ, Austin CP, Inglese J (2009) A basis for reduced chemical library inhibition of firefly luciferase obtained from directed evolution. J Med Chem 52:1450–1458
Thorne N, Shen M, Lea WA, Simeonov A, Lovell S, Auld DS, Inglese J (2012) Firefly luciferase in chemical biology: a compendium of inhibitors, mechanistic evaluation of chemotypes, and suggested use as a reporter. Chem Biol 19:1060–1072
Dranchak P, MacArthur R, Guha R, Zuercher WJ, Drewry DH, Auld DS, Inglese J (2013) Profile of the GSK published protein kinase inhibitor set across ATP-dependent and-independent luciferases: implications for reporter-gene assays. PLoS One 8, e57888
Davis MI, Sasaki AT, Shen M, Emerling BM et al (2013) A homogeneous, high-throughput assay for phosphatidylinositol 5-phosphate 4-kinase with a novel, rapid substrate preparation. PLoS One 8, e54127
Patel PR, Sun H, Li SQ, Shen M, Khan J, Thomas CJ, Davis MI (2013) Identification of potent Yes1 kinase inhibitors using a library screening approach. Bioorg Med Chem Lett 23:4398–4403
Rees MG, Davis MI, Shen M, Titus S, Raimondo A, Barrett A, Gloyn AL, Collins FS, Simeonov A (2014) A panel of diverse assays to interrogate the interaction between glucokinase and glucokinase regulatory protein, two vital proteins in human disease. PLoS One 9, e89335
Inglese J, Auld DS, Jadhav A, Johnson RL, Simeonov A, Yasgar A, Zheng W, Austin CP (2006) Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc Natl Acad Sci U S A 103:11473–11478
Yasgar A, Shinn P, Jadhav A, Auld D, Michael S, Zheng W, Austin CP, Inglese J, Simeonov A (2008) Compound management for quantitative high-throughput screening. JALA Charlottesv Va 13:79–89
Rix U, Hantschel O, Durnberger G, Remsing Rix LL et al (2007) Chemical proteomic profiles of the BCR-ABL inhibitors imatinib, nilotinib, and dasatinib reveal novel kinase and nonkinase targets. Blood 110:4055–4063
Copeland RA (2005) Evaluation of enzyme inhibitors in drug discovery. A guide for medicinal chemists and pharmacologists. Methods Biochem Anal 46:1–265
Jenkins WT (1991) The pyruvate kinase-coupled assay for ATPases: a critical analysis. Anal Biochem 194:136–139
Demian DJ, Clugston SL, Foster MM, Rameh L, Sarkes D, Townson SA, Yang L, Zhang M, Charlton ME (2009) High-throughput, cell-free, liposome-based approach for assessing in vitro activity of lipid kinases. J Biomol Screen 14:838–844
Lloyd DJ, St Jean DJ Jr, Kurzeja RJ, Wahl RC et al (2013) Antidiabetic effects of glucokinase regulatory protein small-molecule disruptors. Nature 504:437–440
Acknowledgement
This work was supported by the Molecular Libraries Initiative of the NIH Roadmap for Medical Research. The content of this publication does not necessarily reflect the views of policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.
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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. In: Zegzouti, H., Goueli, S. (eds) Kinase Screening and Profiling. Methods in Molecular Biology, vol 1360. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3073-9_4
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DOI: https://doi.org/10.1007/978-1-4939-3073-9_4
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