The mechanism of inhibition of antibody-based inhibitors of membrane-type serine protease 1 (MT-SP1)
The mechanism of inhibition of antibody-based inhibitors of membrane-type serine protease 1 (MT-SP1)
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DOI：
10.1016/j.jmb.2007.03.078
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发表时间：
2007-06-15
影响因子：
5.6
通讯作者：
Craik, Charles S.
中科院分区：
文献类型：
--
作者：
Farady, Christopher J.;Sun, Jeonghoon;Craik, Charles S.
两种新型scFv抗体抑制剂对丝氨酸蛋白酶MT-SP1/基质酶的抑制机制揭示了其效力和特异性的基础。动力学实验表明，这些抑制剂具有极强的K-I抑制剂，其值在低皮摩尔范围内，与S1位点的底物结合竞争。对蛋白酶活性位点周围环的丙氨酸扫描为抑制剂特异性提供了理论依据。每个抗体与活性位点两侧的一些残基结合，形成一个独特的三维结合表位。有趣的是，一种抑制剂以类似底物的方式结合在活性位点缝隙中，可以在低pH下被MT-SP1处理，并且是蛋白酶的标准机制抑制剂。这些抑制机制为这些抑制剂的有效性提供了理论依据，并表明开发针对密切相关酶家族个体成员的特异性抗体抑制剂是可行的，并且是开发工具来梳理复杂生物过程的有效方法。(C) 2007 Elsevier Ltd.版权所有。
The mechanisms of inhibition of two novel scFv antibody inhibitors of the serine protease MT-SP1/matriptase reveal the basis of their potency and specificity. Kinetic experiments characterize the inhibitors as extremely potent inhibitors with K-I, values in the low picomolar range that compete with substrate binding in the S1 site. Alanine scanning of the loops surrounding the protease active site provides a rationale for inhibitor specificity. Each antibody binds to a number of residues flanking the active site, forming a unique three-dimensional binding epitope. Interestingly, one inhibitor binds in the active site cleft in a substrate-like manner, can be processed by MT-SP1 at low pH, and is a standard mechanism inhibitor of the protease. The mechanisms of inhibition provide a rationale for the effectiveness of these inhibitors, and suggest that the development of specific antibody-based inhibitors against individual members of closely related enzyme families is feasible, and an effective way to develop tools to tease apart complex biological processes. (C) 2007 Elsevier Ltd. All rights reserved.