Thursday, March 28, 2013

Applications Thursday: High-Throughput MEKK2 Intrinsic ATPase Activity Assay

In the current issue of the Journal of Biomolecular Screening the Scott lab from North Carolina Central published a paper entitled: 'Development and Validation of a High-Throughput Intrinsic ATPase Activity Assay for the Discovery of MEKK2 Inhibitors'. The kinase MEKK2 is implicated in cancer so a selective inhibitor could be a useful therapeutic agent. In the current report the authors exploit the finding that MEKK2 can hydrolyze ATP even when there is no substrate for MEKK2 to phosphorylate.

Following incubation of MEKK2 with ATP the production of ADP could be monitored using the ADP-Glo Reagent from Promega and the resulting luminescence was detected using the PHERAstar microplate reader from BMG LABTECH. Using the PHERAstar they were able to perform detection in a high-throughput assay that had average Z' values of 0.72; indicating that this assay will be reliable for high-throughput screening. An initial screen for inhibitors of MEKK2 ATPase activity using this high-throughput format revealed 2 novel inhibitors.

To find out more about the diverse applications that the PHERAstar and other BMG LABTECH microplate readers can perform please visit the applications center on the BMG LABTECH website.

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