Unraveling the dynamics that enable unusual heme enzyme reactivity

揭示血红素酶异常反应的动力学

• 批准号: 10798604
• 负责人: Katherine Marie Davis
• 金额: $ 8.5万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10798604
• 关键词: Anabolism; Antibiotics; Behavior; Biological; Biotechnology; Cell Respiration; Communicable Diseases; Development; Disease; Enzymes; Goals; Health; Heme; Human; Hydroxylation; Immune response; Immune system; Life; Medicine; Metals; Methodology; Methods; Outcome; Pathway interactions; Pharmacologic Substance; Process; Production; Reaction; Regulation; Research; Role; Signal Transduction; Site-Directed Mutagenesis; System; Therapeutic; Time; Visualization; X-Ray Crystallography; analog; anti-cancer therapeutic; biochemical tools; cancer therapy; catalyst; cofactor; combat; fascinate; hormone biosynthesis; interest; metalloenzyme; nitration; structural biology

ABSTRACT Enzymes are crucial biological catalysts that expedite challenging reactions across all domains of life. While the development of structural biology methods over the past century has enabled visualization of these fascinating systems, our understanding of the dynamic processes that facilitate enzyme reactivity remains limited due to the timescales on which they occur. The overarching goal in my research group is to develop and apply time-resolved methods capable of visualizing these processes to elucidate the mechanisms of metal-containing enzymes important for human health and medicine. Heme-dependent enzymes are of particular interest due to their role in aerobic metabolism ranging from signal transduction, antibiotic biosynthesis and immune response. Despite sharing similar structural motifs and catalytic intermediates, heme-dependent enzymes are capable of catalyzing a wide range of reactions beyond their archetypal hydroxylation outcomes. This project aims to investigate the structural features and dynamic behavior that enables this atypical reactivity from dioxygenation to nitration and beyond. In particular, we plan to repurpose biochemical tools capable of pausing turnover, such as substrate/cofactor analogs and site-directed mutagenesis, as well as apply state-of-the-art time- resolved methods that my group is currently developing, to visualize short-lived catalytic intermediates via a combination of X-ray crystallographic and spectroscopic approaches. Although applied to specific systems herein, the proposed methodologies may have utility in the study of heme-enzymes more broadly, as well as other metalloenzymes. Likewise, the anticipated results have the potential to impact both biocatalysis and the downstream development of biotechnologies and therapeutics in the treatment of cancers and infectious diseases.

抽象的 酶是至关重要的生物催化剂，可以加快所有领域的挑战性反应 生活。尽管过去一个世纪的结构生物学方法的发展已实现 这些引人入胜的系统的可视化，我们对动态过程的理解 由于发生的时间尺度，促进酶反应性仍然有限。这 我的研究小组中的总体目标是开发和应用能够的时间分辨方法 可视化这些过程以阐明重要的含金属酶的机制 用于人类健康和医学。血红素依赖性酶特别感兴趣 在信号转导，抗生素生物合成和免疫的有氧代谢中的作用 回复。尽管共享相似的结构基序和催化中间体，但血红素依赖性 酶能够催化广泛的反应以外的原型 羟基化结果。该项目旨在调查结构特征和动态 从二氧化到硝化及以后的这种非典型反应性的行为。在 特别是，我们计划重新利用能够暂停营业额的生化工具，例如 底物/辅因子类似物和定向诱变，并应用最新的时间 我的小组目前正在开发的解决方法，以可视化短寿命的催化 中间通过X射线晶体学和光谱方法的组合。 尽管本文适用于特定系统，但所提出的方法可能具有实用性 血红素酶的研究更广泛，以及其他金属酶。同样，预期 结果有可能影响生物催化和下游的发展 在癌症和传染病治疗中的生物技术和治疗剂。