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Expert Opin. Drug Metab. Toxicol. 4, 103–120. Bioluminescent assays for ADMET 2008

Cali, J.J., Niles, A., Valley, M.P., O’Brien, M.A., Riss, T.L., and Shultz, J.

Notes: The authors of this review article highlight the use of bioluminescence as a readout for high-throughput ADME/Tox assays. They discuss three strategies for designing bioluminescent assays, using either luciferase, ATP or luciferin substrates as the limiting reagents for a luciferase-catalyzed reaction. Reporter gene assays limit the production of luciferase by tying it to a promoter or DNA regulatory region of interest. Such assays can be used to study genes that are regulated by drugs and other xenobiotics. Bioluminescent assays in which ATP is the limiting reagent of the luciferase reaction can be designed to monitor cell viability or the activity kinases. Bioluminescent assays in which the substrate is limiting can be designed so that the activity of a particular enzyme results in the production of a luciferin substrate that can, in turn, be acted upon by luciferase. (3926)

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J. Biomol. Scr. 13, 870-878. Identification of small molecule inhibitors of β-amyloid cytotoxicity through a cell-based high-throughput screening platform. 2008

Seyb, K.I., Schuman, E.R., Ni, J., Huang, M.M., Michaelis, M.L., and Glicksman, M.A.

Notes: This paper demonstrates use of a calpain assay in a cell-based format. (Calpain-Glo™ Assay). (3941)

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Assay Drug Dev. Technol. 5, 127–136. Bioluminescent assays for high-throughput screening 2007

Fan, F. and Wood, K.V.

Notes: The authors of this paper review bioluminescent assay technologies, discussing HTS reporter, cell-based and luciferase biosensor assays. They divide luminescent assays into three basic categories: assays that measure ATP concentration (cell viability and kinase assays), assays that measure changes in luciferase levels (reporter assays, GPCR assays), and assays that measure changes in luciferin levels (protease [including caspase], P450 and MAO assays). (3737)

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Proc. Natl. Acad. Sci. USA 104, 11103-11108. Chronic exposure to paclitaxel diminishes phosphoinositide signaling by calpain-mediated neuronal calcium sensor-1 degradation. 2007

Boehmerle, W., Zhang, K., Sivula, M., Heidrich, F.M., Lee, Y. Jordt, S-E., and Ehrlich, B.E.

Notes: Taxol-induced peripheral neuropathy is a common side-effect of treatment that has been associated with disturbed intracellular calcium homeostasis in neuronal cells. These authors investigated whether prolonged exposure to Taxol caused alterations in calcium signaling in human neuroblastoma and rat dorsal root ganglia. They found that expression of the inositol 1,4,5-triphosphate receptor modulator NCS-1 was reduced in Taxol-treated neuronal cells. The authors also found that Taxol treatment activated calpain, which degrades NCS-1, and that the calpain inhibitor AK295 prevented Taxol-mediated suppression of calcium release. These findings suggest that calcium-mediated activation of calpain is responsible for the observed degradation of NCS-1. (3681)

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