Kunugi S, Yokoyama M, Sakamoto A
Department of Polymer Science and Engineering, Kyoto Institute of Technology, Japan.
FEBS Lett. 1995 Apr 3;362(2):189-91. doi: 10.1016/0014-5793(95)00242-2.
The pre-steady state process in the thermolysin-catalyzed hydrolysis of Cbz-Gly-Phe-Ala was observed at pH 4.5 by fluorescence stopped-flow method using Dns-Phe as a displacement probe. After the confirmation of the pre-equilibrium hypothesis for the binary interaction, the nonlinear substrate concentration dependence of the apparent kinetic constant for the pre-steady state process was analyzed and an existence of multi-intermediates was proposed.
在pH 4.5条件下，使用丹磺酰苯丙氨酸（Dns-Phe）作为置换探针，通过荧光停流法观察了嗜热菌蛋白酶催化水解苄氧羰基-甘氨酰-苯丙氨酰-丙氨酸（Cbz-Gly-Phe-Ala）的预稳态过程。在确认了二元相互作用的预平衡假设后，分析了预稳态过程表观动力学常数对底物浓度的非线性依赖性，并提出存在多个中间体。
