Membrane Fusion, Organization, and Dynamics Using Supported Bilayers

使用受支持的双层的膜融合、组织和动力学

• 批准号: 7924959
• 负责人: STEVEN G. BOXER
• 金额: $ 9.69万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7924959
• 关键词: Architecture; Binding; Biological; Biological Assay; Biological Models; Biological Process; Biotechnology; Characteristics; Classification; Complementary DNA; Complex; DNA; Deposition; Development; Devices; Diffuse; Diffusion; Docking; Drug Combinations; Elements; Fluorescence; Fluorescence Microscopy; Funding; G-Protein-Coupled Receptors; Gated Ion Channel; Goals; Grant; Health; Human; Image; Incidence; Individual; Integral Membrane Protein; Interferometry; Ion Channel; Isotopes; Kinetics; Lateral; Lead; Lipid Bilayers; Lipids; Liquid substance; Mass Spectrum Analysis; Measurement; Measures; Membrane; Membrane Fluidity; Membrane Fusion; Membrane Lipids; Membrane Potentials; Membrane Proteins; Methods; Microscopy; Monitor; Oligonucleotides; Optics; Pattern; Pattern Formation; Phase; Population; Positioning Attribute; Process; Proteins; Reaction; Reporting; Resolution; SNAP receptor; Simulate; Solid; Structure; Surface; System; Technology; Testing; Time; Vesicle; Work; analytical method; conformational conversion; design; drug development; novel; research study; small molecule; tool; two-dimensional; voltage

DESCRIPTION (provided by applicant): The long-term goals of this project are to develop methods to probe the organization and dynamic reorganization of biological membranes. This includes interactions among the components within membranes, interactions between membrane surfaces that lead to binding, fusion, and pattern formation, and conformational changes of proteins associated with membranes. The lipid bilayer is the basic structure common to biological membranes. Membrane fluidity is critical for biological functions that depend upon conformational changes within membranes, the lateral association or clustering of multiple components, and processes that change membrane topology such as edo- and exocytocis and fusion. This proposal outlines new types of experiments that probe these basic aspects of membrane dynamics using tools that have been developed to pattern, manipulate and image supported bilayers. During the next grant period the focus will be on the mechanism of vesicle fusion, using vesicles that are tethered to supported bilayers and whose interactions can be monitored at the level of individual vesicles (Aim 1); the lateral association and organization of lipids and membrane anchored proteins using a novel type of imaging mass spectrometry that permits membrane composition analysis with unprecedented lateral resolution, sensitivity and information content (Aim 2); and the design and fabrication of an integrated optical/electrical device that will permit high precision interferometry on planar bilayers to probe conformational transitions of membrane-associated proteins, with an initial focus on voltage-gated ion channels (Aim 3). Each aim depends upon the development of new supported lipid bilayer architectures and analytical methods that can have a broad impact on studies of biological membranes. Relevance to human health: A significant fraction of all proteins are associated with membranes, and, as a class, these constitute a huge and diverse target for drug development. This proposal outlines new methods for studying membranes and membrane-associated proteins that can impact our understanding of biological function and organization, as well as impact biotechnology.

描述（由申请人提供）：该项目的长期目标是开发探测生物膜的组织和动态重组的方法。这包括膜内组件之间的相互作用，导致结合，融合和模式形成的膜表面之间的相互作用以及与膜相关的蛋白质的构象变化。脂质双层是生物膜共有的基本结构。膜流动性对于依赖于膜内构象变化，多个成分的横向关联或聚类以及改变膜拓扑的过程（例如Edo-和cococytocis and Fusion）的过程至关重要。该建议概述了使用已开发出用于模式，操纵和图像支持的双层的工具来探测膜动力学的这些基本方面的新型实验类型。在下一个赠款期间，重点将放在囊泡融合的机理上，使用囊泡束缚在支撑的双层囊中，并且可以在单个囊泡的水平上监测其相互作用（AIM 1）；脂质和膜的横向关联和组织使用一种新型的成像质谱法锚定蛋白质，该质谱允许使用前所未有的横向分辨率，灵敏度和信息含量（AIM 2）允许膜组成分析；以及集成的光学/电气设备的设计和制造，该设备将允许对平面双层的高精度干涉法探测与膜相关蛋白的构象转变，并初始关注电压门控离子通道（AIM 3）。每个目标都取决于开发新的受支持的脂质双层架构和分析方法，这些方法可能对生物膜研究产生广泛影响。与人类健康的相关性：所有蛋白质的很大一部分都与膜有关，作为一类，这些蛋白质构成了药物开发的巨大而多样的靶标。该建议概述了研究膜和与膜相关的蛋白质的新方法，这些方法可能会影响我们对生物学功能和组织的理解以及影响生物技术。