Inhibition of human caspases by peptide-based and macromolecular inhibitors
Inhibition of human caspases by peptide-based and macromolecular inhibitors
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DOI：
10.1074/jbc.273.49.32608
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发表时间：
1998-12-04
影响因子：
4.8
通讯作者：
Thornberry, NA
中科院分区：
文献类型：
--
作者：
Garcia-Calvo, M;Peterson, EP;Thornberry, NA
半胱氨酸蛋白酶家族的肽基和大分子抑制剂的研究有助于确定这些酶在炎症和哺乳动物细胞凋亡中的核心作用。由于对这些分子的选择性理解不完全，对这些研究的明确解释受到了损害。在这里，我们描述了几个肽为基础的抑制剂和coxpox丝氨酸蛋白酶抑制剂CrmA对10人半胱天冬酶的选择性。检测的肽醛（Ac-WEHD-CHO、Ac-DEVD-CHO、Ac-YVAD-CHO、叔丁氧羰基-IETD-CHO和叔丁氧羰基-AEVD-CHO）包括几种含有各种半胱天冬酶最佳四肽识别基序的肽醛。这些醛显示出对这些酶的宽范围的选择性和效力，解离常数范围从75 pM到>10 μ M。卤甲基酮苄氧基羰基-VAD氟甲基酮是一种广泛特异性的不可逆半胱天冬酶抑制剂，二级灭活率范围为2.9 x 10（2）M-1 s（-1）（半胱天冬酶-2）至2.8 x 10（5）M-1 s（-1）（半胱天冬酶-1）。基于肽的抑制剂获得的结果与先前描述的底物特异性研究预测的结果雅阁。牛痘丝氨酸蛋白酶抑制剂CrmA是I组半胱天冬酶（半胱天冬酶-1、-4和-5）和大多数III组半胱天冬酶（半胱天冬酶-8、-9和-10）的有效（Ki < 20 nM）和选择性抑制剂，表明该病毒通过抑制细胞凋亡和宿主炎症反应促进感染。
Studies with peptide-based and macromolecular inhibitors of the caspase family of cysteine proteases have helped to define a central role for these enzymes in inflammation and mammalian apoptosis. A clear interpretation of these studies has been compromised by an incomplete understanding of the selectivity of these molecules. Here we describe the selectivity of several peptide-based inhibitors and the coxpox serpin CrmA against 10 human caspases. The peptide aldehydes that were examined (Ac-WEHD-CHO, Ac-DEVD-CHO, Ac-YVAD-CHO, t-butoxycarbonyl-IETD-CHO, and t-butoxycarbonyl-AEVD-CHO) included several that contain the optimal tetrapeptide recognition motif for various caspases. These aldehydes display a wide range of selectivities and potencies against these enzymes, with dissociation constants ranging from 75 pM to >10 mu M. The halomethyl ketone benzyloxycarbonyl-VAD fluoromethyl ketone is a broad specificity irreversible caspase inhibitor, with second-order inactivation rates that range from 2.9 x 10(2) M-1 s(-1) for caspase-2 to 2.8 x 10(5) M-1 s(-1) for caspase-1. The results obtained with peptide-based inhibitors are in accord with those predicted from the substrate specificity studies described earlier. The cowpox serpin CrmA is a potent (K-i < 20 nM) and selective inhibitor of Group I caspases (caspase-1, -4, and -5) and most Group III caspases (caspase-8, -9, and -10), suggesting that this virus facilitates infection through inhibition of both apoptosis and the host inflammatory response.