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Author:
Date:
2015-08-20
[Abstract] Glutamate dehydrogenase (GDH) is an NAD(H) dependent enzyme that catalyzes, in vitro, the reversible amination of glutamate. Here we describe how to determine spectrophotometrically GDH activity monitoring NADH evolution. This protocol is described here for Arabidopsis thaliana (A. thaliana) although it is also valid for other plant species. GDH protein is a hexamer composed, in the case of Arabidopsis, of a combination of GDHα, GDHβ and GDHγ subunits. Every combination of subunits is possible; however, it is still barely known whether different combinations affect the enzymatic properties of the hexamers. In other species, hexamers are a combination of GDHα and GDHβ but it cannot be discarded the existence of other genes since for instance GDHγ subunit ...
[摘要] 谷氨酸脱氢酶（GDH）是NAD（H）依赖性酶，其在体外催化谷氨酸盐的可逆胺化。在这里我们描述如何确定分光光度法GDH活动监测NADH进化。此协议在这里描述为拟南芥（ A thaliana ），虽然它也适用于其他植物物种。 GDH蛋白是在拟南芥属的情况下由GDHα，GDHβ和GDHγ亚基的组合组成的六聚体。每个亚基的组合是可能的;然而，仍然几乎不知道不同的组合是否影响六聚体的酶性质。在其他物种中，六聚体是GDHα和GDHβ的组合，但不能丢弃其它基因的存在，因为例如在Fontaine等人（2012）中描述了拟南芥中的GDHγ亚基。
谷氨酸盐+ NAD + + H + →2-氧代戊二酸+ NADH + NH4 +
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