Dynamically-resolved structural studies of proteins from fungal and bacterial pathogens

真菌和细菌病原体蛋白质的动态解析结构研究

• 批准号: 10711972
• 负责人: Diana Canelhas Freitas Monteiro
• 金额: $ 4.11万
• 依托单位: HAUPTMAN-WOODWARD MEDICAL RESEARCH INST
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-10 至 2023-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10711972
• 关键词: Anabolism; Antifungal Agents; Aromatic Amino Acids; Biochemical; Burkholderia pseudomallei; Candida auris; Catalysis; Collaborations; Communicable Diseases; Dedications; Drug Design; Drug Targeting; Drug resistance; Enzymes; Fungal Proteins; Grant; Knowledge; Laboratories; Medical; Methodology; Methods; Organic Synthesis; Oxidative Stress; Pharmaceutical Chemistry; Pharmaceutical Preparations; Protein Conformation; Proteins; Regulation; Research; Sampling; Structure; Technology; Temperature; Therapeutic; Time; Visualization; Work; biological adaptation to stress; biophysical analysis; combat; desensitization; design; drug discovery; experimental study; inorganic phosphate; insight; multidisciplinary; novel; novel therapeutics; pathogen; pathogenic bacteria; pathogenic fungus; programs; protein function; protein structure; structural biology

Summary Drug resistance and desensitization is a growing global concern. To overcome these therapeutic issues, drug discovery pipelines have to make use of all available structural information to generate novel, potent and safe therapeutics. Current medicinal chemistry efforts typically make use of static, cryo-temperature protein structures, which do not integrate the dynamics known govern protein function. Advancing from structure-activity to dynamic-activity relationships will diversify the way we pursue drug discovery, opening doors to more advanced therapeutics. To obtain this knowledge, the projects proposed will pursue time-resolved structural studies of medically-relevant proteins from fungal and bacterial pathogens. Although challenging, time- resolved structural biology has matured over the last decade making it amenable to a wider range of protein targets. This research will break methodological barriers to swiftly obtain dynamic- activity relationships, from which we will determine how dynamics correlate to protein function. Complementary biochemical and biophysical studies will be used to unambiguously resolve protein metastable intermediates. Methods such as photocaging and cryo-trapping are used to trigger synchronize protein function in the sample, making use of the work and expertise of the PI as well as the available facilities, including a dedicated organic synthesis laboratory. Two initial targets will be pursued. The first one, C. Auris 3-Deoxy-D-arabinoheptulosonate 7- phosphate Synthase (DAHPS) is an enzyme essential for fungal survival as it catalysis a step in the biosynthesis of aromatic amino-acids. It is considered a promising drug target to current antifungals and it is dynamically allosterically regulated. The second target is rubrerythrin from B. pseudomallei, a protein believed to be responsible for oxidative stress responses for which the mechanism is still unclear. Both projects will be targeted using an overarching methodological pipeline which will generate new insights into dynamic-activity relationships and provide structures of short-lived intermediates. Beyond the cutting-edge work proposed in this grant, these discoveries will fuel a long-term research program focusing on harnessing dynamic information to drive novel drug design. The fundamental work proposed in this grant will fill gaps in our knowledge of protein mechanism and its relationship to dynamics, and will pave the way to novel drug discovery approaches. The proposed time-resolved experiments are challenging but made possible by the multi-disciplinary expertise of the PI as well as strong collaborations that drive fertile scientific discussions and unobstructed access to cutting edge technology and facilities in the USA and abroad.

总结 耐药性和脱敏是一个日益严重的全球问题。为了克服这些治疗 药物发现管道必须利用所有可用的结构信息， 产生新的、有效的和安全的疗法。目前的药物化学工作通常使 使用静态的低温蛋白质结构，其不整合已知的动力学 控制蛋白质的功能。从结构-活动关系到动态-活动关系， 使我们寻求药物发现的方式多样化，为更先进的治疗方法打开大门。到 获得这方面的知识，拟议的项目将进行时间分辨结构研究， 来自真菌和细菌病原体的医学相关蛋白质。虽然时间很长，但... 解析结构生物学在过去十年中已经成熟，使其适用于更广泛的领域。 蛋白质目标。这项研究将打破方法上的障碍，迅速获得动态- 活性关系，从中我们将确定动力学如何与蛋白质功能相关。 补充的生物化学和生物物理研究将用于明确解决 蛋白质亚稳态中间体。使用诸如光笼化和低温捕获的方法来 利用PI的工作和专业知识，触发同步样本中的蛋白质功能 以及现有的设施，包括一个专门的有机合成实验室。 将追求两个初步目标。第一个，C。7-脱氧-D-阿拉伯庚酮糖酸耳 磷酸合酶（DAHPS）是真菌生存所必需的酶，因为它催化一个步骤， 芳香族氨基酸的生物合成。它被认为是一个有前途的药物靶点， 抗真菌药，并且它是动态变构调节的。第二个靶标是来自B的赤藓红菌素。 pseudomallei，一种被认为是负责氧化应激反应的蛋白质， 机制尚不清楚。这两个项目都将采用一种总体方法 管道，这将产生对动态活动关系的新见解，并提供结构 短暂的中间体。除了这项资助中提出的尖端工作外， 这些发现将推动一项长期研究计划，重点是利用动态信息 to drive驱动novel新drug药物design设计. 这项资助所提出的基础性工作将填补我们在蛋白质机制方面的知识空白 及其与动力学的关系，并将为新药发现方法铺平道路。的 提出的时间分辨实验是具有挑战性的，但由于多学科 PI的专业知识以及强有力的合作推动了丰富的科学讨论， 畅通无阻地获得美国和国外的尖端技术和设施。