From poor substrates to good inhibitors: Design of inhibitors for serine and thiol proteases
From poor substrates to good inhibitors: Design of inhibitors for serine and thiol proteases
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DOI：
10.1021/bi952879e
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发表时间：
1996-03-19
期刊：
影响因子：
2.9
通讯作者：
Abeles, RH
中科院分区：
文献类型：
--
作者：
Baggio, R;Shi, YQ;Abeles, RH
丝氨酸和巯基蛋白酶与肽底物反应形成酰基酶。我们已经合成了抑制剂，它们是假底物，与蛋白酶反应产生缓慢水解的酰基酶。这通过在抑制剂I [Ac-Phe-C（O）NH-NH-C（O）X]和II [苄基-O-C（O）-Psi-Ala-Leu-ArgOMe]的羰基附近引入给电子基团来实现。来自I与木瓜蛋白酶和II与胰凝乳蛋白酶的反应的酰基酶水解锡的12和1小时，分别。抑制剂羰基上电子密度的增加也降低了酰基酶形成的速率。将与离去基团结合位点（S'亚位点）相互作用并加速抑制剂与酶的反应速率的组分掺入抑制剂中。对于抑制剂I，X = NH（CH 3），与木瓜蛋白酶反应的k（on）< 0.13 M（-1）s（-1），但如果X = Psi Leu（CH 3）（2），k（on）= 10（5）M（-1）s（-1）。用II和胰凝乳蛋白酶获得了类似的结果。伴随着酰基酶的形成，X被释放，缓慢水解的酰基酶保留下来。
Serine and thiol proteases react with peptide substrates to form an acyl-enzyme. We have synthesized inhibitors which are pseudo-substrates and react with the proteases to generate acyl-enzymes which hydrolyze slowly. This is achieved by incorporating an electron-donating group near the carbonyl group of inhibitors I [Ac-Phe-C(O)NH-NH-C(O)X] and II [benzyl-O-C(O)-Psi-Ala-Leu-ArgOMe]. The acyl-enzymes derived from the reaction of I with papain and II with chymotrypsin hydrolyze with tin of 12 and 1 h, respectively. The increased electron density on the carbonyl group of the inhibitor also reduces the rate of acyl-enzyme formation. Components were incorporated into the inhibitor which interact with the leaving group binding site (S' subsite) and which accelerate the rate of reaction of inhibitor with enzyme. For inhibitor I, X = NH(CH3), k(on) < 0.13 M(-1) s(-1) for the reaction with papain, but if X = Psi Leu(CH3)(2), k(on) = 10(5) M(-1) s(-1). Similar results were obtained with II and chymotrypsin. Concomitant with acyl-enzyme formation, X is released and a slowly hydrolyzing acyl-enzyme remains.