人线粒体ClpP蛋白酶变构活化的机制

IF 9.1 1区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Monica M. Goncalves, Adwaith B. Uday, Taylor J. B. Forrester, S. Quinn W. Currie, Angelina S. Kim, Yue Feng, Yulia Jitkova, Algirdas Velyvis, Robert W. Harkness, Matthew S. Kimber, Aaron D. Schimmer, Natalie Zeytuni, Siavash Vahidi
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引用次数: 0

摘要

人ClpP蛋白酶通过降解错误折叠蛋白参与线粒体蛋白质量控制。ClpP在急性髓性白血病(AML)等癌症中过度表达,其抑制导致受损呼吸链亚基的积累和细胞死亡。相反,用小分子激活剂(如最近发现的ONC201)过度激活ClpP,会破坏线粒体蛋白的降解,损害癌细胞的呼吸。尽管它在人类健康中起着至关重要的作用,但人类ClpP的结构和功能特性背后的机制仍然难以捉摸。值得注意的是,人类ClpP被活性位点抑制剂矛盾地激活。迄今为止发表的所有可用的人类ClpP结构都处于非活性紧密或压缩状态,即使ClpP与ONC201等激活剂分子结合也令人惊讶。在这里,我们展示了处于活性延伸状态的人类线粒体ClpP的结构,包括一对ClpP与活性位点抑制剂结合的结构。我们发现,把手区(A192E和E196R)的氨基酸替换重建了细菌ClpP中发现的保守盐桥,稳定了扩展活性状态,显著提高了ClpP活性。我们阐明了ClpP的激活机制,强调了一种激发效应,其中亚化学计量抑制剂结合触发变构转变,驱动ClpP进入其活性扩展状态。我们的研究结果将ClpP的构象动力学与其催化功能联系起来,并为合理设计有效和特异性的ClpP抑制剂提供了高分辨率结构,这对靶向AML和其他与ClpP有关的疾病具有重要意义。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Mechanism of allosteric activation in human mitochondrial ClpP protease
Human ClpP protease contributes to mitochondrial protein quality control by degrading misfolded proteins. ClpP is overexpressed in cancers such as acute myeloid leukemia (AML), where its inhibition leads to the accumulation of damaged respiratory chain subunits and cell death. Conversely, hyperactivating ClpP with small-molecule activators, such as the recently discovered ONC201, disrupts mitochondrial protein degradation and impairs respiration in cancer cells. Despite its critical role in human health, the mechanism underlying the structural and functional properties of human ClpP remains elusive. Notably, human ClpP is paradoxically activated by active-site inhibitors. All available structures of human ClpP published to date are in the inactive compact or compressed states, surprisingly even when ClpP is bound to an activator molecule such as ONC201. Here, we present structures of human mitochondrial ClpP in the active extended state, including a pair of structures where ClpP is bound to an active-site inhibitor. We demonstrate that amino acid substitutions in the handle region (A192E and E196R) recreate a conserved salt bridge found in bacterial ClpP, stabilizing the extended active state and significantly enhancing ClpP activity. We elucidate the ClpP activation mechanism, highlighting a hormetic effect where substoichiometric inhibitor binding triggers an allosteric transition that drives ClpP into its active extended state. Our findings link the conformational dynamics of ClpP to its catalytic function and provide high-resolution structures for the rational design of potent and specific ClpP inhibitors, with implications for targeting AML and other disorders with ClpP involvement.
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来源期刊
CiteScore
19.00
自引率
0.90%
发文量
3575
审稿时长
2.5 months
期刊介绍: The Proceedings of the National Academy of Sciences (PNAS), a peer-reviewed journal of the National Academy of Sciences (NAS), serves as an authoritative source for high-impact, original research across the biological, physical, and social sciences. With a global scope, the journal welcomes submissions from researchers worldwide, making it an inclusive platform for advancing scientific knowledge.
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