Structural and mechanistic insights into antibiotic resistance in Staphylococcus aureus

金黄色葡萄球菌抗生素耐药性的结构和机制见解

• 批准号: 2272004
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2272004
• 关键词: Structural; mechanistic; insights; into; antibiotic

Background: Antibiotic resistance (AR) undermines effective antibacterial chemotherapy and constitutes a major threat to global public health. A comprehensive response to this problem includes gaining a detailed understanding of the mechanisms by which AR is mediated. This project seeks to characterize the structural and molecular mechanism of several antibiotic resistance proteins found in the 'superbug', Staphylococcus aureus; resistance systems that are not currently well understood. In the first instance, we will focus on uncharacterized determinants capable of mediating resistance the aminoglycosides or mupirocin. Objectives: To elucidate, with the aid of structural characterization, the molecular mechanism by which staphylococcal resistance proteins mediate resistance to clinically-important antibiotics acting on the bacterial translation machinery.Novelty: Despite their adverse impact on the clinical use of antibiotics, no structural or mechanistic detail currently exists for the resistance proteins that will be studied here. Such information will be crucial to inform the development of strategies to overcome or circumvent these resistance mechanisms.Timeliness: Antibiotic resistance is a major global concern at present, considered by the WHO to be one of the greatest threats to human health, and features prominently in the funding priorities of the research councils, including the BBSRC. The structural aspects of the study will employ cryo-EM, and will therefore make use of the state-of-the art facilities recently established at Leeds.Experimental approach: For both initial targets of interest we already have access to purified, soluble protein. For the larger of the two, crystallography has already proven unsuccessful, and we will therefore use cryo-EM for structural characterization. For the smaller protein, we have in preliminary studies been able to generate crystals that require optimization - hence, the initial effort in this case will focus on obtaining an X-ray structure. The structural insights gained in these studies will inform downstream mutagenesis and biochemical experiments to interrogate function.

背景：抗生素耐药(AR)破坏了有效的抗菌化疗，并构成对全球公共健康的主要威胁。对这个问题的全面回应包括对AR的调节机制有一个详细的了解。该项目旨在表征在“超级细菌”--金黄色葡萄球菌中发现的几种抗生素耐药蛋白的结构和分子机制；这些耐药系统目前尚不清楚。在第一个例子中，我们将专注于能够调节氨基糖苷或莫匹罗星耐药性的未知决定因素。目的：借助结构特征，阐明葡萄球菌耐药蛋白介导临床重要抗生素耐药作用于细菌转译机的分子机制。新奇的是：尽管葡萄球菌耐药蛋白对抗生素的临床应用有不利影响，但目前尚无结构或机制细节。这些信息将对制定克服或绕过这些耐药机制的战略至关重要。及时性：抗生素耐药性是目前全球关注的主要问题，被世卫组织视为对人类健康的最大威胁之一，并在包括BBSRC在内的研究委员会的资助优先事项中占有突出地位。这项研究的结构方面将使用低温EM，因此将利用利兹最近建立的最先进的设施。实验方法：对于这两个感兴趣的初始目标，我们已经可以获得纯化的、可溶的蛋白质。对于两者中较大的一个，结晶学已经被证明是不成功的，因此我们将使用低温电子显微镜进行结构表征。对于较小的蛋白质，我们在初步研究中已经能够产生需要优化的晶体-因此，在这种情况下，最初的努力将集中在获得X射线结构上。从这些研究中获得的结构洞察力将为下游突变和生物化学实验提供信息，以询问功能。