PES inhibits human-inducible Hsp70 by covalent targeting of cysteine residues in the substrate-binding domain.
PES inhibits human-inducible Hsp70 by covalent targeting of cysteine residues in the substrate-binding domain.
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PES 通过共价靶向底物结合域中的半胱氨酸残基抑制人诱导型 Hsp70
DOI：
10.1074/jbc.ra120.015440
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Perrett S
中科院分区：
文献类型：
--
作者：
Yang J;Gong W;Wu S;Zhang H;Perrett S
Hsp70 蛋白是一个古老且保守的伴侣蛋白家族。它们在重要的细胞过程中发挥着重要作用，例如蛋白质质量控制和应激反应。 Hsp70 蛋白是治疗疾病，特别是癌症的潜在药物靶点。据报道，PES（2-苯乙炔磺酰胺或 Pifithrin-μ）是 Hsp70 的抑制剂。然而，PES抑制的机制仍不清楚。在这项研究中，我们发现 PES 可以与人 HspA1A (hHsp70) 的 SBDα 中的 Cys-574 和 Cys-603 发生迈克尔加成反应，导致 PES 分子与每个 Cys 残基共价连接。我们之前表明 Cys-574 和 Cys-603 的谷胱甘肽化影响 hHsp70 的结构和功能。在这项研究中，PES 修饰对 hHsp70 表现出与谷胱甘肽类似的结构和功能影响。此外，我们发现对 PES 修饰的敏感性受到 SBDα 构象动力学变化的影响，例如由与相邻结构域的相互作用、变构变化和突变引起的。该研究为hHsp70共价抑制剂的开发提供了新途径。
Hsp70 proteins are a family of ancient and conserved chaperones. They play important roles in vital cellular processes, such as protein quality control and the stress response. Hsp70 proteins are a potential drug target for treatment of disease, particularly cancer. PES (2-phenylethynesulfonamide or pifithrin-μ) has been reported to be an inhibitor of Hsp70. However, the mechanism of PES inhibition is still unclear. In this study we found that PES can undergo a Michael addition reaction with Cys-574 and Cys-603 in the SBDα of human HspA1A (hHsp70), resulting in covalent attachment of a PES molecule to each Cys residue. We previously showed that glutathionylation of Cys-574 and Cys-603 affects the structure and function of hHsp70. In this study, PES modification showed similar structural and functional effects on hHsp70 to glutathionylation. Further, we found that susceptibility to PES modification is influenced by changes in the conformational dynamics of the SBDα, such as are induced by interaction with adjacent domains, allosteric changes, and mutations. This study provides new avenues for development of covalent inhibitors of hHsp70.