Structural Biology of Oxylipin Biosynthesis

氧脂质生物合成的结构生物学

基本信息

批准号：
7579832
负责人：
MICHAEL G MALKOWSKI
金额：
$ 33.83万
依托单位：
HAUPTMAN-WOODWARD MEDICAL RESEARCH INST
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-02-01 至 2012-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7579832
关键词：
Acetylation Active Sites Adverse effects Affect Amino Acids Anabolism Animals Anti-Inflammatory Agents Anti-inflammatory Arachidonic Acids Arthritis Aspirin Binding Biological Carbon Cardiovascular system Caribbean region Catalysis Chemicals Chemistry Clinical Complex Crystallization Cytochromes Data Detergents Development Dioxygenases Disease Eicosanoids Enzymes Exhibits Family Fatty Acids Food Generations Goals Health Homeostasis Homology Modeling Human Hydroxyeicosatetraenoic Acids Immune response Inflammatory Invertebrates Investigation Leukotrienes Life Style Linoleic Acids Lipids Lipoxins Lipoxygenase Mediator of activation protein Membrane Methods Modeling Molecular Molecular Conformation Mus Mutagenesis Mutation Non-Steroidal Anti-Inflammatory Agents Oryza sativa Oxygen Oxygenases Perception Pharmacia brand of valdecoxib Physiological Processes Plants Play Polyunsaturated Fatty Acids Process Production Property Prostaglandin H2 Prostaglandin Production Prostaglandin-Endoperoxide Synthase Prostaglandins Proteins Reaction Renal function Reproduction Research Personnel Resolution Rofecoxib Role Site-Directed Mutagenesis Specificity Structure System Testing Therapeutic base carboxylate celecoxib coral cyclooxygenase 1 cyclooxygenase 2 inhibitor/antagonist insight interest lipid mediator mutant novel pathogen programs protoporphyrin IX stereochemistry structural biology vascular inflammation

项目摘要

DESCRIPTION (provided by applicant): OBJECTIVE: The broader goal of this proposal is to understand how different integral membrane enzymes utilize stereoselective oxygenations to generate unique oxylipins from a defined set of polyunsaturated fatty acid (PUFA) substrates. Oxylipins are bioactive lipid mediators that are biosynthesized from 18-22 carbon PUFAs through the addition of molecular oxygen via the catalytic activity of cytochrome P450s, lipoxygenases, and cyclooxygenases (COX-1 and COX-2). One of the most biologically important groups of oxylipins is the eicosanoid class, which include prostaglandins (PGs) and leukotrienes derived from arachidonic acid. These products are responsible for the modulation of basic physiologic processes and act as potent lipid mediators of the inflammatory process and other immune responses. COX-1 and COX-2 are the pharmacological target of aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs), including the COX-2 specific inhibitors Vioxx, Bextra, and Celebrex. SPECIFIC AIMS: Using mutagenesis, functional analyses, and x-ray crystallographic methods, we will (1) elucidate the structure of pathogen-inducible oxygenase and characterize at the molecular level the mechanism and structural determinants involved in the stereoselective oxygenation of 18 carbon PUFAs into 2R-oxylipin products; and (2) elucidate the structures of unique 15R-PG producing COX:PUFA complexes in order to understand at the molecular level how the conformation of the PUFA in the active site influences stereospecific oxygenation in the generation of these novel products. HEALTH RELEVANCE: The role that PUFAs play in health and disease is generating renewed interest, with a more focused public perception of healthy food and lifestyle and the significant impact that these compounds have in certain clinical conditions. The ability of proteins, such as COX-2, to dramatically shift their product profiles upon treatment with pharmacological inhibitors has led to further investigations into how these enzymes function. Our studies will provide for a complete mechanistic understanding of how novel lipids are derived from PUFAs upon aspirin treatment, and lend valuable insight into development of new or combined therapeutic approaches for the management of arthritis and vascular inflammation, with fewer unwanted side effects.

描述（由申请人提供）：目的：该提案的更广泛的目标是了解不同的积分膜酶如何利用立体选择性氧合来从一组定义的多不饱和脂肪酸（PUFA）底物中产生独特的氧基蛋白。阿氧蛋白是生物活性脂质介质，通过通过细胞色素P450S，脂肪氧合酶和环氧酶（COX-1和COX-2）的催化活性（COX-1和COX-2）添加分子氧，从18-22碳PUFA生物合成。黄霉素最重要的oxylipins是类固醇类别，其中包括源自蛛网膜酸的前列腺素（PGS）和白细胞。这些产品负责调节基本生理过程，并充当炎症过程和其他免疫反应的有效脂质介质。 COX-1和COX-2是阿司匹林和其他非甾体类抗炎药（NSAIDS）的药理靶标，包括COX-2特异性抑制剂Vioxx，Bextra和Celebrex。具体目的：使用诱变，功能分析和X射线晶体学方法，我们将（1）阐明病原体可诱导的氧合酶的结构，并在分子水平表征与18碳氧化的机制和结构决定因素，将18个碳氧化物的定型性氧化剂纳入2R-氧化脂蛋白产物；（2）阐明产生独特的15R-PG的结构：PUFA复合物，以便在分子级别理解活性位点中PUFA的构象如何影响这些新产品的产生的立体特异性氧合。健康相关性：PUFA在健康和疾病中所起的作用引起了新的兴趣，更加集中于公众对健康食品和生活方式的认识以及这些化合物对某些临床状况的重大影响。蛋白质（例如COX-2）在用药理学抑制剂治疗后急剧转移其产品曲线的能力已导致进一步研究这些酶的功能。我们的研究将提供对阿司匹林治疗后如何从pufas衍生出的新型脂质的完整机械理解，并为开发用于治疗关节炎和血管炎症的新型或联合治疗方法的有价值的见解，副作用较少。