Université Paris-Saclay, CNRS, INRAe, Université Paris Cité, Université d'Evry, Institute of Plant Sciences Paris-Saclay (IPS2), Gif-sur-Yvette, France.
Department of Plant Biochemistry, Albrecht-von-Haller Institute for Plant Sciences, University of Gottingen, Gottingen, Germany.
Methods Mol Biol. 2024;2792:29-39. doi: 10.1007/978-1-0716-3802-6_3.
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 O and releases HO. The oxidation of o-dianisidine by HO 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.
磷酸甘油酸磷酸酶 (PGLP) 将 2-磷酸甘油酸去磷酸化生成甘油酸，甘油酸可以进一步被氧化酶 (GOX) 代谢为乙醛酸，通过氧化反应消耗 O 并释放 HO。HO 催化的邻茴香胺的氧化可以通过在 440nm 处的吸光度变化实时跟踪。基于这些反应，描述了一种使用与重组拟南芥 GOX 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 PGLP 或从拟南芥莲座叶中提取的总可溶性蛋白。
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