A method for identification of inhibitors of the phosphorylation reactionsof bacterial response regulator proteins using P-31 nuclear magnetic resonance spectroscopy

Bacterial response regulators are attractive targets for antibacterial drug development, yet random screening against these targets has failed as yet to identify chemicals that constitute viable leads. Alternative methods to provide leads for drug development based on identification and optimization of low affinity ligands from NMR screens have been described. However, lea ds from these processes still require verification in a bioassay, which is often problematic if compounds have unfavorable optical and solubility prop erties. A simple method, based on using NMR to observe the activity of the target, is described. It has the advantages of being able to characterize b oth low affinity leads and a wider selection of compounds in a structure ac tivity relationships series, without the problems affecting a fluorescence assay. In this example we use P-31 to monitor the turnover of a bacterial r esponse regulator, but the generic approach could be applied to other nucle i and thus a range of biological systems. (C) 2001 Elsevier Science.