Abstract：
Phosphoglycolate phosphatase (PGLP) dephosphorylates 2-phosphoglycolate to glycolate that can be further metabolized to glyoxylate by glycolate oxidase (GOX) via an oxidative reaction that uses O2 and releases H2O2. The oxidation of o-dianisidine by H2O2 catalyzed by a peroxidase can be followed in real time by an absorbance change at 440 nm. Based on these reactions, a spectrophotometric method for measuring PGLP activity using a coupled reaction with recombinant Arabidopsis thaliana GOX is described. This protocol has been used successfully with either purified PGLP or total soluble proteins extracted from Arabidopsis rosette leaves.
摘要：
磷酸乙醇酸磷酸酶(PGLP)将2-磷酸乙醇酸去磷酸化为乙醇酸,乙醇酸可以通过使用O2并释放H2O2的氧化反应被乙醇酸氧化酶(GOX)进一步代谢为乙醛酸。过氧化物酶催化的H2O2对邻二茴香胺的氧化可以实时跟踪440nm处的吸光度变化。基于这些反应,描述了使用与重组拟南芥GOX的偶联反应测量PGLP活性的分光光度法。该方案已成功用于纯化的PGLL或从拟南芥莲座叶提取的总可溶性蛋白质。
