一种用于检测丙氨酰 - 转移RNA合成酶对转移RNA进行氨酰化反应的连续分光光度测定法。
A continuous spectrophotometric assay for the aminoacylation of transfer RNA by alanyl-transfer RNA synthetase.
作者信息
Wu M X, Hill K A
机构信息
Department of Biochemistry, Loma Linda University School of Medicine, California 92350.
出版信息
Anal Biochem. 1993 Jun;211(2):320-3. doi: 10.1006/abio.1993.1276.
A continuous spectrophotometric assay is described for the aminoacylation reaction catalyzed by Escherichia coli alanyl-transfer RNA synthetase. The assay is based on coupling the alanyl-tRNA synthetase-dependent formation of AMP to the lactate dehydrogenase oxidation of NADH. Oxidation of NADH, as monitored at 340 nm, is shown to be stoichiometric with the formation of alanyl-tRNA(Ala). This assay will facilitate the rapid accumulation and analysis of kinetic data for alanyl-tRNA synthetase and should be applicable to aminoacyl-tRNA synthetases in general.
本文描述了一种用于检测大肠杆菌丙氨酰 - 转运RNA合成酶催化的氨酰化反应的连续分光光度法。该检测方法基于将丙氨酰 - tRNA合成酶依赖的AMP形成与NADH的乳酸脱氢酶氧化反应偶联。在340nm处监测到的NADH氧化与丙氨酰 - tRNA（Ala）的形成呈化学计量关系。这种检测方法将有助于快速积累和分析丙氨酰 - tRNA合成酶的动力学数据，并且一般应适用于氨酰 - tRNA合成酶。
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