Toolkit for High-Resolution Structure and Dynamics of Functional Lipids

功能性脂质的高分辨率结构和动力学工具包

• 批准号: 9752610
• 负责人: James H. Morrissey
• 金额: $ 95.51万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-13 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9752610
• 关键词: Address; Algorithmic Analysis; Binding; Biological; Biological Process; Biology; Blood coagulation; Cell Adhesion; Cells; Chemicals; Chronic; Communities; Complement; Computing Methodologies; Crystallization; Data; Data Collection; Databases; Disease; Environment; Eukaryotic Cell; Event; Goals; Health; Heart Diseases; Hemostatic function; Human; In Vitro; Inflammation; Ions; Isotope Labeling; Label; Ligands; Lipid Bilayers; Lipid Binding; Lipids; Magic; Malignant Neoplasms; Measures; Membrane; Membrane Biology; Membrane Lipids; Membrane Proteins; Metals; Methods; Microscopy; Molecular; Molecular Conformation; NMR Spectroscopy; Nerve Degeneration; Nuclear Magnetic Resonance; Pathology; Peripheral; Pharmaceutical Preparations; Phospholipids; Play; Preparation; Production; Proteins; Protocols documentation; Reagent; Relaxation; Resolution; Role; Signal Transduction; Site; Spectrum Analysis; Sterols; Stroke; Structural Genes; Structural Protein; Structure; System; Technology; Vacuum; antimicrobial peptide; cost effective; human disease; innovation; metabolomics; mimetics; molecular dynamics; nanoscale; non-Native; novel strategies; programs; protein function; protein structure; restraint; small molecule; solid state nuclear magnetic resonance; structural biology; tool

Project Summary Membrane proteins are abundant in eukaryotic cells and play important roles in a great many biological processes ranging from cell adhesion and recognition to energy production to signaling cascades. Furthermore, membrane proteins make up about 60% of the targets for currently approved drugs, which underscores their relevance to human disease. Although very high resolution structures of a number of membrane proteins have now been solved, we lack methods that will also allow us to resolve the structures of the membrane lipids that interact with membrane-embedded proteins, peripheral membrane proteins and other ligands. This is in spite of the fact that specific membrane lipids play key regulatory roles in biology. We term these “functional lipids” because, in addition to their well-known structural roles in membranes, it is becoming increasingly clear that lipids are effector molecules that modulate and/or directly carry out essential biological functions. An atomic-scale understanding of the interactions carried out by functional lipids is an important unmet goal with direct relevance to human health and disease. Thus, although excellent methods now exist for solving the structures of proteins—including membrane proteins—at very high resolution, the field lacks tools necessary to solve the structures of the lipid part of membranes at high resolution. This ambitious project aims to develop an innovative “toolkit” of high-resolution methods for the scientific community to use in solving the structures of lipids that regulate the biological functions of membranes. Our approach requires synergistic and coordinated efforts throughout: (1) cost-effective, site-specific isotopic labeling of a variety of phospholipids and sterols; (2) assembling labeled lipids into nanoscale lipid bilayer systems together with their biologically relevant ligands; (3) nuclear magnetic resonance (NMR) approaches, principally high-field magic-angle spinning solid-state NMR (SSNMR), to obtain detailed structural information about the lipids interacting with ligands; (4) cutting-edge computational methods employing molecular dynamics (MD) simulations of lipids interacting with ligands in bilayers or bilayer mimetics; and (5) new methods for solving lipid structures by marrying computational NMR and MD approaches to address the unique challenges inherent in interpreting and understanding spectral data obtained from planar bilayers that contain repeating copies of labeled lipids interacting with neighboring lipids in addition to their specific ligands. As our studies progress, we propose to apply this toolkit to exemplary problems in biology, including blood coagulation, antimicrobial peptide action and sterol recognition.

项目概要 膜蛋白在真核细胞中含量丰富，在许多生物体中发挥着重要作用。 从细胞粘附和识别到能量产生再到信号级联的过程。 此外，膜蛋白约占目前批准药物靶标的 60%， 强调它们与人类疾病的相关性。尽管许多结构的分辨率非常高 现在膜蛋白已经被解决了，我们缺乏能够让我们解析膜蛋白结构的方法 与膜嵌入蛋白、外周膜蛋白和其他蛋白相互作用的膜脂 配体。尽管事实上特定的膜脂在生物学中发挥着关键的调节作用。我们术语 这些“功能性脂质”，因为除了它们在膜中众所周知的结构作用之外，它正在变得 越来越清楚的是，脂质是调节和/或直接执行重要生物作用的效应分子。 功能。从原子尺度理解功能性脂质的相互作用非常重要 与人类健康和疾病直接相关的未实现目标。因此，尽管现在存在优秀的方法 以非常高的分辨率解析蛋白质（包括膜蛋白）的结构，该领域缺乏工具 以高分辨率解析膜的脂质部分的结构是必需的。这个雄心勃勃的项目旨在 开发一个高分辨率方法的创新“工具包”，供科学界用来解决 调节膜生物功能的脂质结构。我们的方法需要协同和 协调一致的努力：（1）对各种磷脂进行具有成本效益的位点特异性同位素标记 甾醇； (2) 将标记的脂质与其生物学特性一起组装成纳米级脂质双层系统 相关配体； (3)核磁共振(NMR)方法，主要是高场魔角 旋转固态核磁共振 (SSNMR)，以获得有关脂质相互作用的详细结构信息 配体； (4) 采用脂质分子动力学（MD）模拟的尖端计算方法 与双层或双层模拟物中的配体相互作用； (5) 解析脂质结构的新方法 将计算 NMR 和 MD 方法相结合，解决解释中固有的独特挑战 并理解从包含重复的标记脂质副本的平面双层获得的光谱数据 除了其特定配体之外，还与邻近的脂质相互作用。随着我们研究的进展，我们建议 将此工具包应用于生物学中的典型问题，包括凝血、抗菌肽作用 和甾醇识别。