Sulisobenzone is a potent inhibitor of the global transcription factor Cra

IF 3 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY
Neetu Neetu , Jai Krishna Mahto , Monica Sharma , Madhusudhanarao Katiki , Preeti Dhaka , Partha Roy , Shailly Tomar , Anoop Narayan , Dinesh Yernool , Pravindra Kumar
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引用次数: 0

Abstract

Transcription is carried out by the RNA polymerase and is regulated through a series of interactions with transcription factors. Catabolite activator repressor (Cra), a LacI family transcription factor regulates the virulence gene expression in Enterohaemorrhagic Escherichia coli (EHEC) and thus is a promising drug target for the discovery of antivirulence molecules. Here, we report the crystal structure of the effector molecule binding domain of Cra from E. coli (EcCra) in complex with HEPES molecule. Based on the EcCra-HEPES complex structure, ligand screening was performed that identified sulisobenzone as an potential inhibitor of EcCra. The electrophoretic mobility shift assay (EMSA) and in vitro transcription assay validated the sulisobenzone binding to EcCra. Moreover, the isothermal titration calorimetry (ITC) experiments demonstrated a 40-fold higher binding affinity of sulisobenzone (KD 360 nM) compared to the HEPES molecule. Finally, the sulisobenzone bound EcCra complex crystal structure was determined to elucidate the binding mechanism of sulisobenzone to the effector binding pocket of EcCra. Together, this study suggests that sulisobenzone may be a promising candidate that can be studied and developed as an effective antivirulence agent against EHEC.

硫异苯酮是一种有效的全球转录因子Cra抑制剂。
转录由RNA聚合酶进行,并通过与转录因子的一系列相互作用进行调节。分解代谢激活物阻遏物(Cra)是一种LacI家族转录因子,调节肠出血性大肠杆菌(EHEC)中毒力基因的表达,因此是发现抗毒力分子的一个有前途的药物靶点。在此,我们报道了大肠杆菌Cra(EcCra)与HEPES分子复合的效应分子结合结构域的晶体结构。基于EcCra-HEPES复合物结构,进行了配体筛选,确定磺异苯酮是EcCra的潜在抑制剂。电泳迁移率转移测定法(EMSA)和体外转录测定法验证了磺异苯酮与EcCra的结合。此外,等温滴定量热法(ITC)实验表明,与HEPES分子相比,磺异苯酮(KD 360nM)的结合亲和力高出40倍。最后,确定了磺异苯酮结合的EcCra复合物的晶体结构,以阐明磺异苯甲酮与EcCra效应结合口袋的结合机制。总之,这项研究表明,磺异苯酮可能是一种很有前途的候选药物,可以作为一种有效的抗肠出血性大肠杆菌毒力剂进行研究和开发。
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
Journal of structural biology
Journal of structural biology 生物-生化与分子生物学
CiteScore
6.30
自引率
3.30%
发文量
88
审稿时长
65 days
期刊介绍: Journal of Structural Biology (JSB) has an open access mirror journal, the Journal of Structural Biology: X (JSBX), sharing the same aims and scope, editorial team, submission system and rigorous peer review. Since both journals share the same editorial system, you may submit your manuscript via either journal homepage. You will be prompted during submission (and revision) to choose in which to publish your article. The editors and reviewers are not aware of the choice you made until the article has been published online. JSB and JSBX publish papers dealing with the structural analysis of living material at every level of organization by all methods that lead to an understanding of biological function in terms of molecular and supermolecular structure. Techniques covered include: • Light microscopy including confocal microscopy • All types of electron microscopy • X-ray diffraction • Nuclear magnetic resonance • Scanning force microscopy, scanning probe microscopy, and tunneling microscopy • Digital image processing • Computational insights into structure
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