NMR-based activity assays have been developed to identify and characterize inhibitors of two nucleoside ribohydrolase enzymes. Protocols are provided for initial compound assays at 500 μM and 250 μM, dose-response assays for determining IC50 values, detergent counter screen assays, jump-dilution counter screen assays, and assays in E. coli whole cells.
Stockman, B. J., Kaur, A., Persaud, J. K., Mahmood, M., Thuilot, S. F., Emilcar, M. B., Canestrari, M., Gonzalez, J. A., Auletta, S., Sapojnikov, V., Caravan, W., Muellers, S. N. NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases. J. Vis. Exp. (148), e59928, doi:10.3791/59928 (2019).