Advancing the Measurement of Lipid Transbilayer Exchange

推进脂质跨双层交换的测量

• 批准号: 1402901
• 负责人: John Conboy
• 金额: $ 15.5万
• 依托单位: University of Utah
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-15 至 2016-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1402901&HistoricalAwards=false
• 关键词: Advancing; Measurement; Lipid; Transbilayer; Exchange

A membrane, only two molecules thick, surrounds all cells and is responsible for controlling the passage of materials in and out of the cell in a selective manner. Our current understanding of the structure and dynamics of cellular membranes emerged in the early 1970?s. However, there is still much we do not know about this seemingly simple "shell" which makes life as we know it possible. A central issue in biology is the movement of molecules across the cellular membrane. This "translocation" is important in the infection of living cells by viruses, the functioning of antibiotics, antiseptics and drugs, and the regulation and growth of cells. There have been a number of studies attempting to find out just how this happens. There are many theories, but no conclusions. Using methods developed in the investigator's laboratory, a novel analytical approach was invented to selectively probe lipid translocation and membrane composition in a lipid bilayer (model cell membrane). The goal of the proposed research is to use this new tool to address some of the central issues concerning molecular motion in membranes. Facilitating an interest in chemistry, biology, physics and mathematics is also an integral part of the proposed studies, with efforts underway to create an innovative science and math education facility at the University of Utah, which will reach out to high school students, college freshmen and the general public, to expand their awareness and knowledge of science. With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. John Conboy from the University of Utah to unravel the complex interplay between the movement of lipid species across the cellular membrane and the establishment of lipid compositional asymmetry. A full understanding of the mechanism by which membrane asymmetry is achieved and maintained in cellular systems has not been realized to date; primarily due to the difficulty of studying membrane biophysical phenomena in a non-destructive or non-perturbing fashion. It has been suggested that lipid membrane asymmetry is maintained by unidirectional lipid transporters, in conjunction with a high energetic barrier to translocation which limits the rate at which lipids might spontaneously translocate across the membrane. However, such a putative flippase has yet to be identified, and a growing number of publications demonstrate cases of rapid spontaneous translocation of phospholipids. The connection between lipid compositional asymmetry and flip-flop in planar supported lipid bilayers will be explored using a novel application of sum-frequency vibrational spectroscopy (SFVS) developed by the PI to selectively probe the asymmetry in a planar-supported lipid bilayer (PSLB). This new surface analytical method allows for the direct detection of lipid flip-flop without the need for a fluorescent or spin-labeled lipid probe, which can alter the measured translocation rates. The goal of this research is to use this surface analytical tool to address some of the central issues concerning the transbilayer movement and establishment of lipid asymmetry in bilayer systems. In addition, the coupling of lipid flip-flop energetics to the establishment of lipid asymmetry will also be explored. These studies are aimed at providing physical insight into the mechanism of lipid compositional asymmetry.

只有两个分子厚的膜包围着所有的细胞，并负责以选择性的方式控制物质进出细胞。 我们目前对细胞膜的结构和动力学的理解出现在1970年初？S. 然而，对于这个看似简单的“外壳”，我们仍然有很多不了解，因为它使我们所知的生命成为可能。 生物学的一个中心问题是分子在细胞膜上的运动。 这种“易位”在病毒感染活细胞、抗生素、防腐剂和药物的功能以及细胞的调节和生长中很重要。 有许多研究试图找出这是如何发生的。 有很多理论，但没有结论。 使用研究者实验室开发的方法，发明了一种新的分析方法来选择性地探测脂质双层（模型细胞膜）中的脂质易位和膜组成。 拟议研究的目标是使用这种新工具来解决有关膜中分子运动的一些核心问题。 促进对化学，生物学，物理学和数学的兴趣也是拟议研究的一个组成部分，正在努力在犹他州大学创建一个创新的科学和数学教育设施，该设施将覆盖高中生，大学新生和公众，以扩大他们的科学意识和知识。有了这个奖项，化学部的生命过程化学计划正在资助犹他州大学的John Conboy博士，以解开脂质物质穿过细胞膜的运动与脂质组成不对称性的建立之间复杂的相互作用。 迄今为止，尚未完全理解细胞系统中膜不对称性的实现和维持机制;主要是由于难以以非破坏性或非扰动方式研究膜生物物理现象。 有人认为，脂质膜的不对称性是由单向脂质转运蛋白，结合高能量的障碍易位，限制了脂质可能自发地跨膜易位的速率。 然而，这样一个假定的翻转酶尚未被确定，越来越多的出版物证明的情况下，迅速自发易位的磷脂。脂质组成的不对称性和触发器之间的连接在平面支持的脂质双层将探索使用和频振动光谱（SFVS）的PI开发的一种新的应用程序，选择性地探测平面支持的脂质双层（PSLB）的不对称性。 这种新的表面分析方法允许直接检测脂质触发器，而不需要荧光或自旋标记的脂质探针，这可能会改变测量的易位率。 本研究的目标是使用这种表面分析工具来解决一些关于跨双层运动和建立双层系统中的脂质不对称性的核心问题。 此外，还将探讨脂质翻转能量学与脂质不对称性建立的耦合。 这些研究的目的是提供物理洞察机制的脂质组成不对称。