Investigation of a cryptic ligand binding site on Plasmodium falciparum Hsp90

IF 3 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Christopher R. Mansfield , Elizabeth L. Taggart , Michael E. Chirgwin , Emily R. Derbyshire
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引用次数: 0

Abstract

The molecular chaperone heat shock protein 90 (Hsp90) has an important role in maintaining proteostasis in Plasmodium parasites, the causative agents of malaria, and is of interest as a potential antimalarial drug target. Inhibitors targeting its well-characterized N-terminal ATP-binding site are lethal, but the development of high-affinity binders with selectivity for the Plasmodium over the human homolog has been challenging given the high conservation of this domain. A binding site in the less conserved Hsp90 C-terminus has been reported to interact with nucleotides and inhibitors in other eukaryotic systems, which could offer an alternative route for antimalarial design. Herein, we characterize the potential ligandability of the C-terminus in the Plasmodium falciparum chaperone PfHsp90 with in silico and in vitro methods. We conducted affinity experiments with both a lysine-reactive nucleotide analog and an ATP resin that support a specific interaction between ATP and a C-terminal truncation of PfHsp90. We further explored the nucleotide structural requirements for this interaction with limited proteolysis experiments, which suggest association with ATP, dATP, and ADP, but not AMP or GTP. Lastly, we employed computational analyses and mutagenesis studies to interrogate the molecular basis for the interaction. Our findings provide the foundation for future studies to assess and develop C-terminal Hsp90 inhibitors against Plasmodium parasites.

Abstract Image

恶性疟原虫Hsp90隐性配体结合位点的研究
分子伴侣热休克蛋白90 (Hsp90)在维持疟原虫(疟疾病原体)的蛋白平衡中起着重要作用,是一种潜在的抗疟疾药物靶点。针对其特征明确的n端atp结合位点的抑制剂是致命的,但考虑到该结构域的高度保守性,开发对疟原虫具有选择性的高亲和力结合物一直具有挑战性。据报道,在其他真核系统中,保守程度较低的Hsp90 c端上的一个结合位点可以与核苷酸和抑制剂相互作用,这可能为抗疟药物的设计提供另一种途径。在此,我们用计算机和体外方法表征了恶性疟原虫伴侣蛋白PfHsp90的c端潜在的配位性。我们对赖氨酸反应性核苷酸类似物和ATP树脂进行了亲和实验,这些树脂支持ATP与PfHsp90的c端截断之间的特异性相互作用。我们通过有限的蛋白水解实验进一步探索了这种相互作用的核苷酸结构要求,结果表明与ATP、dATP和ADP有关,但与AMP或GTP无关。最后,我们采用计算分析和诱变研究来询问相互作用的分子基础。本研究结果为进一步研究和开发抗疟原虫c端Hsp90抑制剂提供了基础。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Bioorganic & Medicinal Chemistry
Bioorganic & Medicinal Chemistry 医学-生化与分子生物学
CiteScore
6.80
自引率
2.90%
发文量
413
审稿时长
17 days
期刊介绍: Bioorganic & Medicinal Chemistry provides an international forum for the publication of full original research papers and critical reviews on molecular interactions in key biological targets such as receptors, channels, enzymes, nucleotides, lipids and saccharides. The aim of the journal is to promote a better understanding at the molecular level of life processes, and living organisms, as well as the interaction of these with chemical agents. A special feature will be that colour illustrations will be reproduced at no charge to the author, provided that the Editor agrees that colour is essential to the information content of the illustration in question.
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