원문정보
초록
영어
The activities of NADPH-dependent enzymes are crucial for appropriate functions in metabolism and energy transformation of biological systems. These enzymes thus function as biomarkers of some diseases and metabolic disorder. Although the importance is being increased, current assay systems need high costs of instrumentation, time-consuming multi-step pretreatment, lower stability and weak signal of NADPH. Here we present a fluorometric assay that circumvents many issues of previous methods, because this system simply measured an enhanced fluorescence of NADPH, generated just after incubation with only mBFP(a metagenome derived NADPH-dependent blue fluorescence protein). Using mBFP, we set up an HTS system for screening of NADP(H)-dependent enzymes and then validated through a pilot screen using a small library. The simplicity and speed of this system can be useful for the functional characterization of the related enzyme and inhibitors