利用荧光素酶生物发光体系测定Mg~(2+)-ATP酶结合和水解活性

本文应用LKB公司的ATP测液建立了Mg~(2 )-ATP酶的ATP结合及水解活性的测定方法;利用国产荧光素酶粗品在连串反应体系中建立测定Mg~(2 )-ATP酶结合活性的方法,并与水解活性相比较.对Mg~(2 )-ATP酶的去脂样品,Mg~(2 )-ATP酶与卵磷脂复合物以及微粒体样所做的测定表明,上述两种方法是可靠、简便的,尤其是利用国产荧光素酶粗品建立的ATP结合活性的测定方法,能避免水解对结合活性测定的干扰,刘其它的酶-底物的结合研究有参考价值.

ASSAY OF BINDING AND HYDROLYZING ACTIVITY OF ATP ASES USING FIREFLY BIOLUMINESCENCE

Two new methods have been developed. In the first method, LKB ATP Monitoring Reagent was used to monitor the binding and hydrolyzing activity by means of light decrease. In the second method, crude luciferase that was cota-minated with creatine kinase, was used to measure the binding activity of ATPase in a consecutive reaction system. The binding and/or bydrolyzing activities of microsome, delipided ATPase and delipided ATPase/lecithin complex were inve-rstigated, comparaed with the hydrolyzing activity and determined with Jorgen-sen's method1]. The results show that the two methods are reliable and conven ient in assay of ATPase. Especially the exact binding activities of ATPase Can bedetermined in spite of hydrolysis of ATP by using the second method which is a good tool for study of binding between enzyme and substract.