Halfpenny A P, Brown P R
J Chromatogr. 1985 Dec 11;349(2):275-82. doi: 10.1016/s0021-9673(01)83783-3.
Hypoxanthine-guanine phosphoribosyl transferase (HGPRTase) and purine nucleoside phosphorylase (PNPase) activities were simultaneously determined in erythrocyte lysates, using the reversed-phase mode of high-performance liquid chromatography. Reaction conditions were developed to provide zero-order kinetics for both enzymes. The activities of the individual enzymes were calculated after incubation of cell lysates with the PNPase substrate, inosine. After sufficient hypoxanthine had been formed to saturate the HGPRTase, the co-enzyme phosphoribosylpyrophosphate and co-factor magnesium (Mg2+) were added to the incubation medium. The enzyme activities were calculated by measurement of the decrease in the PNPase substrate, inosine, and the increase in the HGPRTase product, inosine-5'-monophosphate.
采用高效液相色谱的反相模式，同时测定红细胞裂解液中的次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HGPRTase）和嘌呤核苷磷酸化酶（PNPase）活性。开发了反应条件以提供两种酶的零级动力学。在细胞裂解液与PNPase底物肌苷孵育后，计算各酶的活性。在形成足够的次黄嘌呤以饱和HGPRTase后，将辅酶磷酸核糖焦磷酸和辅因子镁（Mg2+）添加到孵育培养基中。通过测量PNPase底物肌苷的减少和HGPRTase产物5'-肌苷单磷酸的增加来计算酶活性。
