Metalloenzyme structure, function, and as targets for neurodegeneration and bacterial pathogenesis

金属酶的结构、功能以及作为神经变性和细菌发病机制的靶标

• 批准号: 10626767
• 负责人: THOMAS L POULOS
• 金额: $ 59.6万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10626767
• 关键词: Address; Anabolism; Antibiotics; Binding; Biological; Biology; Catalysis; Complex; Coupled; Cytochrome P450; Development; Drug Metabolic Detoxication; Drug Targeting; Electron Transport; Enzymes; Goals; Heme; Hydrogen Peroxide; Investigation; Iron; Knowledge; Methicillin Resistance; Methods; Nature; Nerve Degeneration; Neurodegenerative Disorders; Nitric Oxide Synthase; Nitric Oxide Synthetase Inhibitor; Oxidation-Reduction; Pathogenesis; Pathway interactions; Peroxidases; Peroxides; Play; Proteins; Protons; Reaction; Research; Role; Staphylococcus aureus; Structure; System; Testing; Therapeutic; Thinking; Work; Xenobiotics; cofactor; design; drug metabolism; fundamental research; insight; metalloenzyme; oxidation; pathogenic bacteria; protein protein interaction; steroid metabolism; structural biology; therapeutic target

Heme is one of the most diverse and useful iron-containing cofactors in biology. One of the most diverse functions are in oxidative heme enzymes that are designed to store the oxidizing equivalents of H2O2 or O2 in order to carry out biologically useful oxidation reactions. These include detoxification of toxic peroxides and xenobiotics (ie drug metabolism) and the oxidation of small organic compounds in various biosynthetic pathways such as in steroid metabolism and antibiotic biosynthesis. Structural biology has played a critical role in understanding these enzymes and the Poulos lab has focused primarily, but not exclusively, on peroxidases, cytochromes P450, nitric oxide synthase (NOS), and the various auxiliary proteins required for electron transfer. Owing to the transient nature of redox partner complexes, it has been difficult to determine crystal structures which is why there are so few in the PDB. This gap in our knowledge is especially important in P450s where redox partner binding can play a critical role in controlling where the substrate is oxidized in addition to exerting an effector role critical for proton coupled electron transfer. Recent advances in the well known P450cam system has provided specific hypotheses on the structural changes induced by redox partner binding that are required for O2 activation and has resulted in a rethinking of traditional views on how P450s work. This has generated considerable discussion, some quite controversial, but also has stimulated research to test the validity of some of these new ideas. A critical question being addressed is the generality of redox partner effector control in P450 catalysis in addition to the biological basis for why such a level of control is required for some P450s but not others. NOS is a P450 and has provided deeper insights into O2 activation and substrate oxidation. NOS also has proven to be an important therapeutic target in neurodegenerative diseases and in certain pathogenic bacteria like methicillin resistant Staph aureus (MRSA). Together with the Silverman lab at Northwestern, structure-based methods are being applied to the development of highly selective NOS inhibitors. Overall, the various ongoing projects provide a synergistic mix of fundamental research in heme enzyme function with research having clearly defined biomedical relevance.

血红素是生物学中最多样化和最有用的含铁辅助因子之一。之一 最多样化的功能是氧化血红素酶，旨在存储 H2O2 或 O2 的氧化当量，以进行生物有用的氧化 反应。这些包括有毒过氧化物和异生物质（即药物 代谢）和各种生物合成中小有机化合物的氧化 途径，例如类固醇代谢和抗生素生物合成。结构生物学 在理解这些酶方面发挥了关键作用，Poulos 实验室已经 主要但不限于过氧化物酶、细胞色素 P450、一氧化氮 合成酶（NOS），以及电子转移所需的各种辅助蛋白。欠 由于氧化还原伙伴配合物的瞬态性质，很难确定 晶体结构，这就是 PDB 中晶体结构如此之少的原因。我们的知识差距是 在 P450 中尤其重要，其中氧化还原伙伴结合可以在 除了发挥关键效应子作用外，还控制底物被氧化的位置 用于质子耦合电子转移。众所周知的 P450cam 的最新进展 系统对氧化还原引起的结构变化提供了具体的假设 O2 激活所需的伴侣结合，并导致重新思考 关于 P450 工作原理的传统观点。这引起了广泛的讨论，一些 相当有争议，但也刺激了研究来测试其中一些的有效性 新想法。正在解决的一个关键问题是氧化还原伙伴效应器的通用性 P450 催化中的控制以及为什么达到如此水平的控制的生物学基础 某些 P450 需要，但其他 P450 则不需要。 NOS 是 P450，提供了更深的 对 O2 活化和底物氧化的见解。 NOS 也被证明是 神经退行性疾病和某些致病性疾病的重要治疗靶点 细菌，如耐甲氧西林金黄色葡萄球菌 (MRSA)。与西尔弗曼实验室一起 在西北大学，基于结构的方法正在应用于开发 高选择性NOS抑制剂。总体而言，各种正在进行的项目提供了 血红素酶功能基础研究与以下研究的协同组合 明确定义的生物医学相关性。

项目成果

Proton Relay Network in the Bacterial P450s: CYP101A1 and CYP101D1.

• DOI: 10.1021/acs.biochem.0c00329
• 发表时间: 2020-08-11
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Amaya JA;Batabyal D;Poulos TL
• 通讯作者: Poulos TL

Updating the Paradigm: Redox Partner Binding and Conformational Dynamics in Cytochromes P450.

• DOI: 10.1021/acs.accounts.1c00632
• 发表时间: 2022-02-01
• 期刊: Accounts of chemical research
• 影响因子: 18.3
• 作者: Poulos TL;Follmer AH
• 通讯作者: Follmer AH

Correction to Improvement of Cell Permeability of Human Neuronal Nitric Oxide Synthase Inhibitors Using Potent and Selective 2-Aminopyridine-Based Scaffolds with a Fluorobenzene Linker.

使用带有氟苯连接体的有效且选择性的基于 2-氨基吡啶的支架来校正人神经元一氧化氮合酶抑制剂的细胞渗透性的改善。

• DOI: 10.1021/acs.jmedchem.8b01866
• 发表时间: 2019
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Do,HaT;Wang,Heng-Yen;Li,Huiying;Chreifi,Georges;Poulos,ThomasL;Silverman,RichardB
• 通讯作者: Silverman,RichardB

Partial Opening of Cytochrome P450cam (CYP101A1) Is Driven by Allostery and Putidaredoxin Binding.

• DOI: 10.1021/acs.biochem.1c00406
• 发表时间: 2021-10-05
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Skinner SP;Follmer AH;Ubbink M;Poulos TL;Houwing-Duistermaat JJ;Paci E
• 通讯作者: Paci E

Correction to Nitrile in the Hole: Discovery of a Small Auxiliary Pocket in Neuronal Nitric Oxide Synthase Leading to the Development of Potent and Selective 2-Aminoquinoline Inhibitors.

孔中腈的校正：神经元一氧化氮合酶中小辅助口袋的发现导致了有效和选择性 2-氨基喹啉抑制剂的开发。

• DOI: 10.1021/acs.jmedchem.8b02033
• 发表时间: 2019
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Cinelli,MarisA;Li,Huiying;Chreifi,Georges;Poulos,ThomasL;Silverman,RichardB
• 通讯作者: Silverman,RichardB