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: 10670417
• 负责人: Katherine Marie Davis
• 金额: $ 36.85万
• 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/10670417
• 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，可以加速所有领域的挑战性反应 的生活。虽然过去一个世纪结构生物学方法的发展使得 这些令人着迷的系统的可视化，我们对动态过程的理解 由于其发生的时间尺度，促进酶反应性仍然受到限制。这 我的研究小组的首要目标是开发和应用能够 可视化这些过程以阐明含金属酶的机制很重要 为了人类健康和医学。血红素依赖性酶由于其 在有氧代谢中的作用，包括信号转导、抗生素生物合成和免疫 回复。尽管具有相似的结构基序和催化中间体，但血红素依赖性 酶能够催化超出其原型的广泛反应 羟基化结果。该项目旨在研究结构特征和动态 这种行为使得这种非典型反应能够从双氧化到硝化等等。在 特别是，我们计划重新利用能够暂停周转的生化工具，例如 底物/辅因子类似物和定点诱变，以及应用最先进的时间- 解决了我的小组目前正在开发的方法，以可视化短暂的催化作用 通过 X 射线晶体学和光谱方法相结合的中间体。 尽管本文应用于特定系统，但是所提出的方法可以在以下方面有用： 更广泛地研究血红素酶以及其他金属酶。同样，预期的 结果有可能影响生物催化和下游发展 治疗癌症和传染病的生物技术和疗法。