Latt S A, Auld D S, Valee B L
Proc Natl Acad Sci U S A. 1970 Nov;67(3):1383-9. doi: 10.1073/pnas.67.3.1383.
An approach is described for the simultaneous measurement of catalysis, the distance from an active-site moiety to fluorescent substrate moieties of an enzyme substrate complex, and the microenvironment of such substrate groups. It is illustrated by means of a three-component energy transfer relay system, consisting of cobalt carboxypeptidase, its fluorescent dansylated peptide substrates, and tryptophanyl residues of the enzyme. The mode of procedure can be applied generally to yield conjoint information on functional and structural aspects of active centers of enzymes.
本文描述了一种同时测量催化作用、酶-底物复合物活性位点部分与荧光底物部分之间的距离以及此类底物基团微环境的方法。通过由钴羧肽酶、其荧光丹磺酰化肽底物和该酶的色氨酸残基组成的三组分能量转移中继系统进行了说明。该程序模式可普遍应用,以提供有关酶活性中心功能和结构方面的联合信息。