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分光光度法测定光呼吸谷氨酸:乙醛酸和丝氨酸:乙醛酸转氨酶反应。

Spectrophotometric Assays for Measuring Photorespiratory Glutamate:Glyoxylate and Serine:Glyoxylate Aminotransferase Reactions.

机构信息

Biology Department, Eastern Michigan University, Ypsilanti, MI, USA.

出版信息

Methods Mol Biol. 2024;2792:41-49. doi: 10.1007/978-1-0716-3802-6_4.

Abstract

Glutamate:glyoxylate aminotransferase (GGAT; EC 2.6.1.4) and serine:glyoxylate aminotransferase activities (SGAT; EC 2.6.1.45) are central photorespiratory reactions within plant peroxisomes. Both enzymatic reactions convert glyoxylate, a product of glycolate oxidase, to glycine, a substrate of the mitochondrial glycine decarboxylase complex. The GGAT reaction uses glutamate as an amino group donor and also produces α-ketoglutarate, which is recycled to glutamate in plastids by ferredoxin-dependent glutamate synthase. Using serine, a product of mitochondrial serine hydroxymethyltransferase, as an amino group donor, the SGAT reaction also produces hydroxypyruvate, a substrate of hydroxypyruvate reductase. The activities of these photorespiratory aminotransferases can be measured using indirect, coupled, spectrophotometric assays, detailed herein.

摘要

谷氨酸

乙醛酸氨基转移酶(GGAT;EC 2.6.1.4)和丝氨酸:乙醛酸氨基转移酶(SGAT;EC 2.6.1.45)是植物过氧化物酶体中光合作用的核心反应。这两种酶促反应将乙醛酸(乙醇酸氧化酶的产物)转化为甘氨酸,甘氨酸是线粒体甘氨酸脱羧酶复合物的底物。GGAT 反应使用谷氨酸作为氨基供体,并产生α-酮戊二酸,该物质在质体中通过依赖铁氧还蛋白的谷氨酸合酶循环回谷氨酸。使用丝氨酸(线粒体丝氨酸羟甲基转移酶的产物)作为氨基供体,SGAT 反应还产生了羟丙酮酸,它是羟丙酮酸还原酶的底物。这些光合作用氨基转移酶的活性可以使用间接、偶联、分光光度法测定,本文详细介绍了该方法。

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