L-TRANS-EPOXYSUCCINYL-LEUCYLAMIDO(4-GUANIDINO)BUTANE (E-64) AND ITS ANALOGS AS INHIBITORS OF CYSTEINE PROTEINASES INCLUDING CATHEPSINS B, H AND L
L-TRANS-EPOXYSUCCINYL-LEUCYLAMIDO(4-GUANIDINO)BUTANE (E-64) AND ITS ANALOGS AS INHIBITORS OF CYSTEINE PROTEINASES INCLUDING CATHEPSINS B, H AND L
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DOI：
10.1042/bj2010189
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发表时间：
1982-01-01
影响因子：
4.1
通讯作者：
HANADA, K
中科院分区：
文献类型：
--
作者：
BARRETT, AJ;KEMBHAVI, AA;HANADA, K
0.5 mM浓度的l-反式环氧琥珀酰亮氨酸（4-胍）丁烷（E-64）对丝氨酸蛋白酶、血浆钾化酶、白细胞弹性酶、金属蛋白酶热溶酶、梭菌胶原酶无影响；10 .mu。M-E-64快速灭活半胱氨酸蛋白酶组织蛋白酶B、H、L和木瓜蛋白酶（t0.5[半衰期]= 0.1 ~ 17.3 s）。链球菌的半胱氨酸蛋白酶反应要慢得多，并且clostripain没有不可逆的失活。半胱氨酸依赖性外肽酶二肽基肽酶I被E-64缓慢灭活。组织蛋白酶B和木瓜蛋白酶与E-64相互作用的活性位点定向性质是通过在可逆竞争性抑制剂leeptin存在下对酶的保护以及与D化合物相反的L抑制的立体特异性来建立的。与木瓜蛋白酶相关的半胱氨酸蛋白酶的快速化学计量反应可用于测定酶溶液的操作摩尔浓度和校准速率测定。报道了一系列E-64结构类似物对人组织蛋白酶B、H和大鼠组织蛋白酶L失活的二级表观速率常数，并与其他一些半胱氨酸蛋白酶活性位点抑制剂的速率常数进行了比较。L-反式环氧琥珀酰基乙酰氨基（3-甲基）丁烷对组织蛋白酶B和L的抑制作用强于E-64。富马酰基乙酰氨基（3-甲基）丁烷的反应性比相应的环氧化物低100倍，但与碘乙酸相当。
L-trans-Epoxysuccinyl-leucylamido(4-guanidino)butane (E-64) at a concentration of 0.5 mM had no effect on the serine proteinases plasma kallikrein and leukocyte elastase or the metalloproteinases thermolysin and clostridial collagenase; 10 .mu.M-E-64 rapidly inactivated the cysteine proteinases cathepsins B, H and L and papain (t0.5 [half-life] = 0.1-17.3 s). The streptococcal cysteine proteinase reacted much more slowly, and there was no irreversible inactivation of clostripain. The cysteine-dependent exopeptidase dipeptidyl peptidase I was very slowly inactivated by E-64. The active-site-directed nature of the interaction of cathepsin B and papain with E-64 was established by protection of the enzyme in the presence of the reversible competitive inhibitor leupeptin and by the stereospecificity for inhibition by the L as opposed to the D compound. The rapid stoichiometric reaction of the cysteine proteinases related to papain can be used to determine the operational molarity of solutions of the enzymes and calibrate rate assays. The apparent 2nd-order rate constants for the inactivation of human cathepsins B and H and rat cathepsin L by a series of structural analogs of E-64 are reported and compared with those for some other active-site-directed inhibitors of cysteine proteinases. L-trans-Epoxysuccinlyleucylamido(3-methyl)butane inhibited cathepsins B and L more rapidly than E-64. Fumarylleucylamido(3-methyl)butane was 100-fold less reactive than the corresponding epoxide, but was about as effective as iodoacetate.