Holzman T F, Baldwin T O
Biochemistry. 1981 Sep 15;20(19):5524-8. doi: 10.1021/bi00522a027.
We have found a new class of inhibitors of the bacterial bioluminescence reaction, the N,N-diphenylalkylamines and acids. We have studied the action of one of these compounds 2,2-diphenylpropylamine. The amine was competitive with the long-chain aliphatic aldehyde substrate (Ki congruent to 0.1 mM) but caused an increase in the affinity of the enzyme for reduced riboflavin 5'-phosphate (FMNH2). The inhibitor was attached to Sepharose 6B by a bis(oxirane) spacer, and the interactions of bacterial luciferase with the immobilized ligand were analyzed. The binding of luciferase to the immobilized inhibitor was enhanced by FMNH2 and was decreased by decanal. The results of these studies showed that the 2,2-diphenylpropylamine-luciferase complex has an increased affinity for FMNH2. Likewise, the FMNH2-luciferase complex has an increased affinity for 2,2-diphenylpropylamine. The inhibitor also binds to the enzyme-4a-peroxydihydroflavin complex to block the binding of the aldehyde substrate, while binding of the aldehyde substrate to either the free enzyme or the enzyme-4a-peroxydihydroflavin complex blocks binding of 2,2-diphenylpropylamine.
我们发现了一类新型的细菌生物发光反应抑制剂，即N,N-二苯基烷基胺和酸。我们研究了其中一种化合物2,2-二苯基丙胺的作用。该胺与长链脂肪醛底物具有竞争性（Ki约为0.1 mM），但会导致酶对还原型核黄素5'-磷酸（FMNH2）的亲和力增加。通过双（环氧乙烷）间隔基将抑制剂连接到琼脂糖6B上，并分析了细菌荧光素酶与固定化配体的相互作用。荧光素酶与固定化抑制剂的结合因FMNH2而增强，因癸醛而减弱。这些研究结果表明，2,2-二苯基丙胺-荧光素酶复合物对FMNH2的亲和力增加。同样，FMNH2-荧光素酶复合物对2,2-二苯基丙胺的亲和力也增加。该抑制剂还与酶-4a-过氧二氢黄素复合物结合，以阻断醛底物的结合，而醛底物与游离酶或酶-4a-过氧二氢黄素复合物的结合则会阻断2,2-二苯基丙胺的结合。
