苯酯是酪蛋白水解蛋白酶 P 的强效抑制剂，并揭示了去寡聚化的立体开关,Journal of the American Chemical Society

当前位置： X-MOL 学术 › J. Am. Chem. Soc. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

苯酯是酪蛋白水解蛋白酶 P 的强效抑制剂，并揭示了去寡聚化的立体开关
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2015-06-29 , DOI: 10.1021/jacs.5b03084
Mathias W. Hackl ₁ , Markus Lakemeyer ₁ , Maria Dahmen ₁ , Manuel Glaser ₂ , Axel Pahl ₁ , Katrin Lorenz-Baath ₁ , Thomas Menzel ₁ , Sonja Sievers ₃ , Thomas Böttcher ₄ , Iris Antes ₂ , Herbert Waldmann _{3,

5} , Stephan A. Sieber ₁

Affiliation

酪蛋白水解蛋白酶 P (ClpP) 是一种参与细胞稳态和致病性的中心细菌降解机制。ClpP 的功能作用已经通过基因敲除和β-内酯的使用进行了研究，β-内酯仍然是迄今为止发现的唯一的 ClpP 特异性抑制剂。β-内酯已作为化学工具在多种生物体中操纵 ClpP；然而，它们的效力、选择性和稳定性是有限的。尽管对 ClpP 活性位点的组成和构象灵活性有详细的结构见解，但迄今为止还没有报道设计特定的非 β-内酯抑制剂的合理努力。在这项工作中，对超过 137 000 种化合物的无偏见筛选用于鉴定五种苯酯化合物作为高效的 ClpP 抑制剂，这些抑制剂对细菌、但不是人类 ClpP。在 ClpP 肽酶和蛋白酶抑制试验中，苯基酯的效力大大超过了 β-内酯，并在活的金黄色葡萄球菌细胞中显示出独特的靶标选择性。分析研究表明，虽然苯基酯像天然肽底物一样被裂解，但它们仍然作为酰基酶中间体共价捕获在活性位点中。36 种衍生物的合成和随后的构效关系 (SAR) 研究提供了对对抑制效力和酰化反应性很重要的保守结构元件的见解。此外，酯的 α 位甲基取代基的立体化学类似于肽底物中的氨基酸侧链，影响 ClpP 复合物的稳定性，导致解离成七聚体或保留十四聚体状态。

"点击查看英文标题和摘要"

Phenyl Esters Are Potent Inhibitors of Caseinolytic Protease P and Reveal a Stereogenic Switch for Deoligomerization

Caseinolytic protease P (ClpP) represents a central bacterial degradation machinery that is involved in cell homeostasis and pathogenicity. The functional role of ClpP has been studied by genetic knockouts and through the use of beta-lactones, which remain the only specific inhibitors of ClpP discovered to date. Beta-lactones have served as chemical tools to manipulate ClpP in several organisms; however, their potency, selectivity and stability is limited. Despite detailed structural insights into the composition and conformational flexibility of the ClpP active site, no rational efforts to design specific non-beta-lactone inhibitors have been reported to date. In this work, an unbiased screen of more than 137 000 compounds was used to identify five phenyl ester compounds as highly potent ClpP inhibitors that were selective for bacterial, but not human ClpP. The potency of phenyl esters largely exceeded that of beta-lactones in ClpP peptidase and protease inhibition assays and displayed unique target selectivity in living S. aureus cells. Analytical studies revealed that while phenyl esters are cleaved like native peptide substrates, they remain covalently trapped as acyl-enzyme intermediates in the active site. The synthesis of 36 derivatives and subsequent structure-activity relationship (SAR) studies provided insights into conserved structural elements that are important for inhibition potency and acylation reactivity. Moreover, the stereochemistry of a methyl-substituent at the alpha position to the ester, resembling amino acid side chains in peptide substrates, impacted ClpP complex stability, causing either dissociation into heptamers or retention of the tetradecameric state. Mechanistic insights into this intriguing stereo switch and the phenyl ester binding mode were obtained by molecular docking experiments.

更新日期：2015-06-29

点击分享查看原文

点击收藏

阅读更多本刊新发论文本刊介绍/投稿指南