Houston B, Nimmo H G
Biochem J. 1984 Dec 1;224(2):437-43. doi: 10.1042/bj2240437.
A new purification procedure for rat liver ATP citrate lyase is described. The method reproducibly gives homogenous undegraded enzyme. Steady-state kinetic analysis of ATP citrate lyase was complicated by the presence of ADP, a product of the reaction, in solutions of ATP. The kinetic patterns observed were dependent on whether ADP was removed by the assay system. When assays were performed with a method in which ADP was removed, the results showed that the enzyme obeys a double-displacement mechanism with a phosphoenzyme intermediate. This resolves a controversy between the results of previous kinetic studies and those of isotope-exchange and enzyme-labelling experiments.
本文描述了一种新的大鼠肝脏ATP柠檬酸裂解酶纯化方法。该方法可重复性地得到均一、未降解的酶。ATP柠檬酸裂解酶的稳态动力学分析因反应产物ADP存在于ATP溶液中而变得复杂。观察到的动力学模式取决于ADP是否被检测系统去除。当采用去除ADP的方法进行检测时，结果表明该酶遵循具有磷酸化酶中间体的双置换机制。这解决了先前动力学研究结果与同位素交换和酶标记实验结果之间的争议。
