"Suspended" Bilayers: New Technology to Study the Dynamics of Membrane Structure

描述(由申请人提供)：这是这项挑战奖助金的前提，通过开发可靠的方法来制造被水溶液包围的“悬浮”双层，将极大地促进对膜结构和功能的动力学的研究。本申请涉及挑战区域06-GM-104，膜结构和功能的动力学。目前的技术是由附着在固体表面的“支撑的”双层膜组成的。它们本质上是非动态的，因为它们所含的蛋白质本质上是不能移动的，而且附着的双层是不能变形的。建议的研究分为一系列特定的目标，每个目标对应于产生功能支撑双层的不同电位方法(S)，我们将并行探索，以查看哪一种方法(S)满足使用功能读数如侧向扩散、单分子水平的结合反应和膜动力学(如依赖陷阱的囊泡融合)建立强大的夹层平台所需的标准。通过直接攻击膜结构和功能动力学夹层研究的最关键的技术限制，并成功地生产出包含功能蛋白质的平面无支撑双层膜，我们可以迎接NIGMS(挑战主题06-GM-104)概述的挑战，并启动许多实验室在膜动力学领域的各种见解。这是一个在两年内实现的现实目标，这将对膜研究产生广泛影响，如果没有刺激资金，这就不会发生。 与公共健康相关：在拟议的项目中，我们的目标是产生独立的(悬浮的)功能平面双层，用于膜动力学的研究。现有的技术支撑在固体表面，导致蛋白质不能移动且不能变形的双层。通过攻击这一限制，我们可以迅速启动许多实验室在膜动力学领域的各种见解。

DESCRIPTION (provided by applicant): It is the premise of this Challenge Grant that investigations of the dynamics of membrane structure and function would be tremendously facilitated by the development of robust methods for the manufacture of "suspended" bilayers that are surrounded on both sides by aqueous solutions. This application addresses Challenge Area 06-GM-104, Dynamics of membrane structure and function. Current technology consists of "supported" bilayers that are attached to a solid surface. They are inherently non-dynamic, as proteins they contain are generally immobile by nature, and an attached bilayer is not deformable. The proposed research is organized into a series of specific aims, each corresponding to different potential method(s) for generating functional supported bilayers, which we will explore in parallel, to see which one(s) meet the required criteria for a robust mezzanine platform using functional read-outs like lateral diffusion, binding reactions at the single molecule level, and membrane dynamics as exemplified by SNARE-dependent vesicle fusion. By attacking directly the single most critical technological limitation of mezzanine studies of the dynamics of membrane structure and function, and succeeding in producing planar unsupported bilayers containing functional proteins, we can meet the challenge outlined by NIGMS (Challenge Topic 06-GM-104), and jump start a great variety of insights from many laboratories in the field of membrane dynamics. This is a realistic goal within a two year frame, which will have a broad impact on membrane research, and which would not occur without the stimulus funding. PUBLIC HEALTH RELEVANCE: In the proposed project, we aim to generate freestanding (suspended) functional planar bi-layers for the study of membrane dynamics. Existing technology supported on solid surfaces, results in bi-layers where proteins are immobile and which cannot be deformed. By attacking this limitation, we can jump start a great variety of insights from many laboratories in the field of membrane dynamics.