Monitoring of phosphatase and kinase activity using 31P NMR spectroscopy

Abstract

Traditionally, enzymatic assays require the addition of labels or exogenous supplementation sources. Phosphorus-31 nuclear magnetic resonance (³¹P NMR) spectroscopy has proven to be a powerful and reliable tool, but it is scarcely employed to evaluate enzymatic activity. Herein, we provide a versatile method to directly characterize phosphatase and kinase activities by exploiting the apparently differential ³¹P NMR shifts between substrates and products, without requiring tags or additional reagents. The utility of this method for inhibitor identification is also presented.