KINETIC STUDIES OF ENZYME MECHANISM

这个项目的目的是研究……的过渡状态 用同位素研究卡那霉素核苷酸转移酶的反应 效果。KNT催化来自ATP的核苷酸残基的转移， GTP或UTP结合到卡那霉素A的4？-羟基，失活 氨基糖苷本身。将使用远程标记法来研究 过渡状态。主要和次要影响都将是 学习。首先要确定的是速率限制 步骤是反应的化学步骤，第二个步骤给出一个 原子间过渡态键级变化的测量 没有直接参与卵裂的。因此，标记三磷酸腺苷的合成 对分子和标记类似物进行了研究。间-硝基苄基 三磷酸是三磷酸腺苷的类似物，它将用于同位素 测量。

The purpose of this project is to study the transition state of the reaction of kanamycin nucleotidyltransferase (KNT) using isotope effects. KNT catalyzes the transfer of a nucleotide residue from ATP, GTP or UTP to the 4?-hydroxyl group of kanamycin A, inactivating the aminoglycoside itself. The remote label method will be used to study the transition state. Both primary and secondary effects will be studied. The first ones in order to determine that the rate limiting step is the chemical step of the reaction, the second ones give a measure of the change in bond order in the trasition state in atoms not directly involved in cleavage. Thus, synthesis of labeled ATP molecules and of labeled analog are carried out. m-nitrobenzyl triphosphate is an analog of ATP and it will be used in the isotopic measurements.