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

描述（由申请人提供）：这是这个挑战补助金的前提，膜结构和功能的动态研究将大大促进了强大的方法，用于制造“悬浮”双层，两侧包围水溶液的发展。本申请解决挑战领域06-GM-104，膜结构和功能的动力学。目前的技术由附着在固体表面上的"支撑"双层组成。它们本质上是非动态的，因为它们所包含的蛋白质通常本质上是固定的，并且附着的双层是不可变形的。所提出的研究被组织成一系列特定的目标，每个目标对应于用于生成功能性支撑双层的不同潜在方法，我们将并行探索这些方法，以查看哪些方法符合使用功能性读出的稳健夹层平台所需的标准，如横向扩散，单分子水平的结合反应和膜动力学，如SNARE依赖性囊泡融合。通过直接攻击膜结构和功能动力学的夹层研究的最关键的技术限制，并成功地生产含有功能蛋白质的平面无支撑双层，我们可以满足NIGMS（挑战主题06-GM-104）概述的挑战，并从许多实验室在膜动力学领域的各种见解开始跳跃。这是一个现实的目标，在两年的框架内，这将对膜研究产生广泛的影响，这将不会发生没有刺激资金。 公共卫生相关性：在拟议的项目中，我们的目标是产生独立的（悬浮）功能平面双层膜动力学的研究。现有的技术支持在固体表面上，导致蛋白质不动并且不能变形的双层。通过攻击这一限制，我们可以从膜动力学领域的许多实验室中获得各种各样的见解。

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.