Reversible Inhibitors Arrest ClpP in a Defined Conformational State that Can Be Revoked by ClpX Association.
Reversible Inhibitors Arrest ClpP in a Defined Conformational State that Can Be Revoked by ClpX Association.
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DOI：
10.1002/anie.201507266
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发表时间：
2015-12
影响因子：
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通讯作者：
A. Pahl;Markus Lakemeyer;Marie-Theres Vielberg;Mathias W. Hackl;Jan Vomacka;V. Korotkov;M. Stein
中科院分区：
文献类型：
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作者：
A. Pahl;Markus Lakemeyer;Marie-Theres Vielberg;Mathias W. Hackl;Jan Vomacka;V. Korotkov;M. Stein
酪蛋白溶解蛋白酶P（Caseinolytic protease P，ClpP）是金黄色葡萄球菌致病的重要调节因子。ClpP肽酶活性抑制剂的高通量筛选导致鉴定了该酶类的第一个非共价结合剂。小分子与S.金黄色葡萄球菌ClpP揭示了一种新的结合模式：由于保守残基脯氨酸125的旋转，ClpP被锁定在确定的构象状态，这导致催化三联体的扭曲和肽酶活性的抑制。基于这些结构见解，通过合理设计和虚拟筛选优化了分子，导致衍生物超过了先前ClpP抑制剂的效力。引人注目的是，构象锁被ClpX的结合所推翻，ClpX是一种相关的分子伴侣，其能够通过ClpXP复合物中的底物解折叠进行蛋白水解。因此，相关分子伴侣调节抑制剂结合是一种对于ClpP药物开发重要的意想不到的机制。
Caseinolytic protease P (ClpP) is an important regulator of Staphylococcus aureus pathogenesis. A high-throughput screening for inhibitors of ClpP peptidase activity led to the identification of the first non-covalent binder for this enzyme class. Co-crystallization of the small molecule with S. aureus ClpP revealed a novel binding mode: Because of the rotation of the conserved residue proline 125, ClpP is locked in a defined conformational state, which results in distortion of the catalytic triad and inhibition of the peptidase activity. Based on these structural insights, the molecule was optimized by rational design and virtual screening, resulting in derivatives exceeding the potency of previous ClpP inhibitors. Strikingly, the conformational lock is overturned by binding of ClpX, an associated chaperone that enables proteolysis by substrate unfolding in the ClpXP complex. Thus, regulation of inhibitor binding by associated chaperones is an unexpected mechanism important for ClpP drug development.