Structures and Mechanisms of Iron-Sulfur Proteins in Redox Control and Stress Response

铁硫蛋白在氧化还原控制和应激反应中的结构和机制

• 批准号: 10582482
• 负责人: Limei Zhang
• 金额: $ 20.55万
• 依托单位: UNIVERSITY OF NEBRASKA LINCOLN
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582482
• 关键词: 3-Dimensional; Address; Administrative Supplement; Anabolism; Award; Bacterial Infections; Cell Survival; Cell physiology; Cryoelectron Microscopy; Diabetes Mellitus; Disease; Disease Progression; Funding; Genetic Transcription; Genus Mycobacterium; Goals; Health; Homeostasis; Human; Infection; Iron; Iron-Sulfur Proteins; Ligands; Link; Malignant Neoplasms; Mediating; Molecular; Molecular Sieve Chromatography; Oxidation-Reduction; Parents; Physiological; Play; Preparation; Protein Family; Proteins; Regulation; Research; Research Personnel; Role; Sampling; Stress; Structural Biochemistry; Structure; Sulfhydryl Compounds; Sulfur; System; Transcriptional Regulation; X-Ray Crystallography; biological adaptation to stress; cofactor; design; insight; instrument; light scattering; novel therapeutic intervention; parent project; particle; pathogen; pathogenic bacteria; repaired

PROJECT SUMMARY Iron-sulfur (Fe-S) clusters are ancient cofactors composed of multiple iron and sulfur atoms. Owing to the rich, tunable redox reactivity and selectivity of the cluster, Fe-S proteins play multifaced roles in redox control under both physiological and stress conditions, which is required for maintaining normal cellular functions and cell survival. Thus, Fe-S proteins are tightly linked to health and diseases (such as cancer, diabetes, and bacterial infection) in humans. The parent Maximizing Investigators’ Research Award (MIRA; R35 GM138157) project addresses the critical gaps in structural and mechanistic understanding of Fe-S proteins in the cellular control of redox homeostasis, including: i) the structural basis of redox reactivity and ligand selectivity in Fe-S clusters; ii) the mechanism by which the redox state and integrity of the Fe-S cluster allows these proteins to sense redox states and regulate transcription; iii) the structural biochemistry of Fe-S cluster biosynthesis and regulation; and iv) the role of non- proteinaceous thiols in modulating Fe-S cluster-mediated redox control. The research goals and approaches of the parent project are demonstrated in the three selected Fe-S protein-mediated mechanisms for redox control and stress response in mycobacteria: i) redox sensing and transcriptional regulation by the WhiB-like (Wbl) family proteins; ii) assembly, transfer, and repair of Fe-S clusters by the sulfur utilization factor (SUF) system; and iii) mycothiol in Fe-S cluster homeostasis. This administrative supplement requests funds for the the acquisition of a size-exclusion chromatography/ multiple-angle light scattering-dynamic light scattering (SEC/MALS-DLS) instrument to identify and characterize how Fe-S proteins interact with their functional partners to regulate redox homeostasis and respond to stress. This instrument will be used to screen and characterize the interactions between Fe-S proteins and the functional partners, and to evaluate and optimize sample preparation for three-dimensional structural characterization by X-ray crystallography and single-particle cryo-electron microscopy. The implementation of this instrument within the scope and experimental workflow of the parent MIRA project is expected to provide unprecedented molecular insights and greatly expedite the mechanistic understanding the Fe-S proteins-mediated cellular control of redox homeostasis.

项目摘要 铁硫原子团簇是由多个铁和硫原子组成的古老的辅因子。由于富人， 可调的氧化还原反应性和选择性的集群，铁硫蛋白发挥多方面的作用，在氧化还原控制下， 生理和应激条件，这是维持正常细胞功能和细胞生长所必需的。 生存因此，Fe-S蛋白与健康和疾病（如癌症、糖尿病和细菌性疾病）紧密相关。 感染）。 最大化研究者研究奖（MIRA; R35 GM 138157）项目解决了关键的 Fe-S蛋白在氧化还原稳态的细胞控制中的结构和机制理解上的差距， 包括：i）Fe-S簇合物中氧化还原反应性和配体选择性的结构基础; ii） Fe-S簇的氧化还原状态和完整性允许这些蛋白质感知氧化还原状态并调节 转录; iii）Fe-S簇生物合成和调节的结构生物化学;以及iv）非- 蛋白质硫醇调节Fe-S簇介导的氧化还原控制。的研究目标和方法 在三个选定的Fe-S蛋白介导的氧化还原控制机制中证明了母项目 分枝杆菌中的氧化还原传感和应激反应：i）WhiB样（Wbl）家族的氧化还原传感和转录调控 ii）通过硫利用因子（SUF）系统组装、转移和修复Fe-S簇;和iii） 真菌硫醇在Fe-S簇内稳态中的作用。 本行政补编要求提供资金，用于购置一台分子排阻色谱仪/ 多角度光散射-动态光散射（SEC/MALS-DLS）仪器来识别和表征 Fe-S蛋白如何与其功能伙伴相互作用以调节氧化还原稳态并对应激作出反应。 该仪器将用于筛选和表征Fe-S蛋白与功能蛋白之间的相互作用。 合作伙伴，并评估和优化三维结构表征的样品制备， X射线晶体学和单粒子低温电子显微镜。本文书的执行， MIRA母项目的范围和实验工作流程预计将提供前所未有的分子生物学技术， 深刻的见解和大大加快机制的理解Fe-S蛋白介导的细胞控制氧化还原 体内平衡