KLR-70：细菌 Hsp70 分子伴侣的新型阳离子抑制剂。,Biochemistry

当前位置： X-MOL 学术 › Biochemistry › 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.)

KLR-70：细菌 Hsp70 分子伴侣的新型阳离子抑制剂。
Biochemistry ( IF 2.9 ) Pub Date : 2020-04-24 , DOI: 10.1021/acs.biochem.0c00320
Matthew D Dalphin ₁ , Andrew J Stangl ₁ , Yue Liu ₁ , Silvia Cavagnero ₁

Affiliation

热休克因子 Hsp70 和其他分子伴侣在新生蛋白质折叠中起着核心作用。然而，阐明复杂细胞环境中个体伴侣所执行的任务一直具有挑战性。实现这一目标的一种策略是在特定分子伴侣抑制剂不存在和存在的情况下监测蛋白质生物发生，然后分析两种条件下的折叠结果。通过这种方式，可以看出感兴趣的陪伴者的作用。然而，伴侣蛋白抑制剂（包括众所周知的富含脯氨酸的抗菌肽）的开发充满了不良的副作用，包括蛋白质表达产量的降低。在这里，我们介绍了 KLR-70，这是一种合理设计的大肠杆菌 Hsp70 分子伴侣（也称为 DnaK）的阳离子抑制剂。KLR-70 是一种 14 个氨基酸的肽，带有天然存在的残基，经过工程改造可与 DnaK 底物结合域相互作用。KLR-70 与 DnaK 的相互作用是对映选择性的，其特点是在缓冲溶液中具有高亲和力。重要的是，KLR-70 不会与 DnaJ 和 GroEL/ES 分子伴侣显着相互作用，并且不会在很宽的浓度范围内改变新生蛋白质的生物合成产量。然而，在复杂的大肠杆菌无细胞环境中观察到 KLR-70 的抗 DnaK 活性有所减弱。有趣的是，d 对映异构体 D-KLR-70 与其全 L KLR-70 对应物不同，它不结合 DnaK 和 DnaJ 分子伴侣，但它强烈抑制翻译。这一结果表明这两种对映异构体（KLR-70 和 D-KLR-70）可以作为分子伴侣结合和翻译的正交抑制剂。总之，KLR-70 是一种新型分子伴侣抑制剂，对细菌 Hsp70 具有高亲和力和选择性，并具有帮助解析 Hsp70 在新生蛋白质折叠中的作用的巨大潜力。

"点击查看英文标题和摘要"

KLR-70: A Novel Cationic Inhibitor of the Bacterial Hsp70 Chaperone.

The heat-shock factor Hsp70 and other molecular chaperones play a central role in nascent protein folding. Elucidating the task performed by individual chaperones within the complex cellular milieu, however, has been challenging. One strategy for addressing this goal has been to monitor protein biogenesis in the absence and presence of inhibitors of a specific chaperone, followed by analysis of folding outcomes under both conditions. In this way, the role of the chaperone of interest can be discerned. However, development of chaperone inhibitors, including well-known proline-rich antimicrobial peptides, has been fraught with undesirable side effects, including decreased protein expression yields. Here, we introduce KLR-70, a rationally designed cationic inhibitor of the Escherichia coli Hsp70 chaperone (also known as DnaK). KLR-70 is a 14-amino acid peptide bearing naturally occurring residues and engineered to interact with the DnaK substrate-binding domain. The interaction of KLR-70 with DnaK is enantioselective and is characterized by high affinity in a buffered solution. Importantly, KLR-70 does not significantly interact with the DnaJ and GroEL/ES chaperones, and it does not alter nascent protein biosynthesis yields across a wide concentration range. Some attenuation of the anti-DnaK activity of KLR-70, however, has been observed in the complex E. coli cell-free environment. Interestingly, the d enantiomer D-KLR-70, unlike its all-L KLR-70 counterpart, does not bind the DnaK and DnaJ chaperones, yet it strongly inhibits translation. This outcome suggests that the two enantiomers (KLR-70 and D-KLR-70) may serve as orthogonal inhibitors of chaperone binding and translation. In summary, KLR-70 is a novel chaperone inhibitor with high affinity and selectivity for bacterial Hsp70 and with considerable potential to help in parsing out the role of Hsp70 in nascent protein folding.

更新日期：2020-04-24

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