Structures and Mechanisms of “Heme-oxygenase-like” Non-heme Di-iron Enzymes that Catalyze Complex N-oxygenation and Olefin-installing C–C-Fragmentation Reactions

催化复杂 N-氧化和烯烃安装 C-C 断裂反应的“类血红素加氧酶”非血红素双铁酶的结构和机制

• 批准号: 10428624
• 负责人: JOSEPH M BOLLINGER
• 金额: $ 32.87万
• 依托单位: PENNSYLVANIA STATE UNIVERSITY, THE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-10 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10428624
• 关键词: Active Sites; Affect; Aldehydes; Alkenes; Amaze; Amino Acids; Antibiotics; Antineoplastic Agents; Aromatic Amines; Back; Binding; Biochemical; Bioinformatics; Biotechnology; Burn injury; Carbon; Chemicals; Clinical; Complex; Coupled; DNA biosynthesis; Data; Dioxygen; Directed Molecular Evolution; Electron Transport; Electron Transport Complex III; Electrons; Enzymes; Equilibrium; Exhibits; Family; Ferredoxin; Ferritin; Goals; Heme; Histidine; Hour; Hydrocarbons; In Situ; In Vitro; Individual; Ions; Iron; Isomerism; Isotopes; Kinetics; Knowledge; Ligands; Malignant neoplasm of pancreas; Mediating; Microbe; Mixed Function Oxygenases; Natural Product Drug; Nitroimidazoles; Oxidants; Oxidases; Oxides; Oxidoreductase; Oxygenases; Pathway interactions; Pharmaceutical Preparations; Plants; Play; Production; Proteins; Publishing; Reaction; Recycling; Reducing Agents; Reporting; Role; Scaffolding Protein; Site; Spectrum Analysis; Streptozocin; Structure; System; Testing; Variant; Work; absorption; abstracting; adduct; carboxylate; cofactor; desaturase; design; enzyme activity; expectation; experimental study; insight; member; microbial; novel; oxidation; scaffold; small molecule

Project Summary/Abstract Enzymes that use coupled di-iron clusters that are coordinated by carboxylate and histidine residues to activate dioxygen for difficult oxidation reactions play crucial roles in the global carbon cycle, in DNA biosynthesis, and in synthesis of clinically used natural- product drugs. In this last year, a new structural family of diiron enzymes has come into focus. Related in structure to heme-oxygenase (HO), these HO-like diiron oxidase and oxygenases (HODOs) have already expanded the known catalytic repertoire of the diiron unit, even with only four members of the new family having been assigned biochemical functions. In their functions, these HO-like enzymes produce antibiotics (the nitroimidazoles), cancer drugs (streptozotocin), and jet fuel. Moreover, they appear to function in a manner that is distinct from the functional paradigm that was established by earlier work on other systems. Rather than remaining as stable cofactors within the HO- proteins scaffolds, they spontaneously degrade, at least in vitro, perhaps as part of a novel modus operandi that eliminates the requirement for cooperating proteins in their catalytic cycles. The goal of this project is to understand the structures and mechanisms of the first four functionally assigned members of what appears, on the basis of bioinformatic analysis, to be a large and versatile new enzyme family. The expectation is that an understanding of its functional principles might enable the new family to become a privileged scaffold for directed evolution of new synthetically useful enzyme activities.

项目摘要/摘要 使用羧酸盐协调和 组氨酸残基激活二氧化物以实现困难的氧化反应在 全球碳循环，在DNA生物合成中，并合成临床使用的自然 产品药物。在去年，一个新的二龙酶家族已经进入 重点。与血红素 - 氧合酶（HO），这些HO样二氧氧化酶和 氧酶（Hodos）已经扩大了二龙的已知催化曲目 单位，即使只有四个新家庭成员被分配了生化 功能。这些HO样酶在其功能中产生抗生素（ 硝基咪唑），癌症药物（链霉菌素）和喷气燃料。而且，他们似乎 功能以与由功能范式不同的方式 早期在其他系统上进行工作。而不是在Ho-中保留为稳定的辅因子 蛋白质支架，它们自发降解，至少在体外，也许是 新颖的作案手法消除了在其中合作蛋白质的要求 催化周期。该项目的目的是了解结构和机制 在前四个功能分配的成员中，出现的成员 生物信息学分析，成为一个大型且多才多艺的新酶家族。期望是 对其功能原则的理解可能使新家庭成为一个 特权脚手架，用于定向新的合成有用酶活性的演变。