KINETIC STUDIES OF ENZYME MECHANISM

本项目的目的是研究 卡那霉素核苷酸转移酶同位素反应 方面的影响. KNT催化从ATP转移核苷酸残基， GTP或UTP到4？-卡那霉素A的羟基，灭活 氨基糖苷本身。 远程标记方法将用于研究 过渡状态。 主要和次要影响都将是 研究了 第一个是为了确定 第一步是反应的化学步骤，第二步是反应的化学步骤， 原子过渡态中键序变化的量度 不直接参与卵裂。 因此，标记ATP的合成 分子和标记的类似物进行。 间硝基苄 三磷酸是ATP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.