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: 9352363
• 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/9352363
• 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; Genes; 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; 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; 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.

项目摘要 膜蛋白在真核细胞中丰富，并且在许多生物学中起重要作用 从细胞粘附和识别到能源产生到信号级联的过程。 此外，膜蛋白约占当前批准药物的60％ 强调了它们与人类疾病的相关性。虽然许多多个分辨率结构 现在已经解决了膜蛋白，我们缺乏可以使我们解决的结构的方法 与膜上包裹的蛋白相互作用的膜脂质，周围膜蛋白和其他 配体。尽管特定的膜脂质在生物学中起关键的调节作用，但这是事实。我们术语 这些“功能性脂质”，因为除了它们在膜中著名的结构作用外， 越来越清楚的是，脂质是调节和/或直接执行必需生物学的效应分子 功能。对功能脂质进行的相互作用的原子级理解是重要的 与人类健康和疾病直接相关的未达到目标。尽管现在存在出色的方法 解决蛋白质的结构（包括膜蛋白），在很高的分辨率下，该领域缺乏工具 在高分辨率下解决膜的脂质部分的结构所必需的。这个雄心勃勃的项目的目标 开发一种创新的高分辨率方法的“工具包”，供科学界用于解决 调节膜的生物学功能的脂质结构。我们的方法需要协同作用和 整个过程：（1）各种磷脂的成本效益，特定地点的同位素标记 （2）将标记的脂质组装到纳米级脂质双层系统及其生物学上 相关配体； （3）核磁共振（NMR）方法，主要是高场魔法角 旋转固态NMR（SSNMR），以获取有关脂质与之相互作用的详细结构信息 配体； （4）采用分子动力学（MD）模拟脂质的尖端计算方法 与双层或双层模拟物中的配体相互作用； （5）通过 嫁给计算NMR和MD方法，以应对解释固有的独特挑战 并了解从包含标记脂质重复副本的平面双层获得的光谱数据 除特定的配体外，还与相邻脂质相互作用。随着我们的研究进展，我们建议 将此工具包应用于生物学的示例性问题，包括血液凝血，抗菌胡椒作用 和固醇识别。