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: 7546649
• 负责人: STEVEN G. BOXER
• 金额: $ 29.04万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2011-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7546649
• 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组织和动态重组的方法。这包括膜内成分之间的相互作用，导致结合、融合和图案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。膜的流动性对于生物功能至关重要，生物功能取决于膜内的构象变化、多个成分的横向关联或聚集，以及改变膜拓扑结构的过程，例如胞吐、胞吐和融合。该提案概述了新型实验，这些实验使用已开发的用于图案化、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。在下一个资助期内，重点将放在囊泡融合机制上，使用与支持的双层相连的囊泡，并且可以在单个囊泡水平上监测其相互作用（目标 1）；使用新型成像质谱法对脂质和膜锚定蛋白进行横向关联和组织，允许以前所未有的横向分辨率、灵敏度和信息内容进行膜成分分析（目标 2）；设计和制造集成光学/电气设备，该设备将允许在平面双层上进行高精度干涉测量，以探测膜相关蛋白的构象转变，最初的重点是电压门控离子通道（目标3）。每个目标都取决于新支持的脂质双层结构和分析方法的开发，这些结构和分析方法可以对生物膜的研究产生广泛的影响。与人类健康的相关性：所有蛋白质中很大一部分与膜相关，作为一类蛋白质，它们构成了药物开发的巨大且多样化的目标。该提案概述了研究膜和膜相关蛋白的新方法，这些方法可以影响我们对生物功能和组织的理解，并影响生物技术。