SPECTROSCOPY ON LENS MEMBRANE LIPID

我们所提议的调查的总体目标是利用 利用最先进的光谱技术 构象;组成和结构特征 透镜膜脂质和蛋白质。 光谱学的发现可以 与膜的组成和性能有关。 我们将 研究正常牛和人类晶状体、白内障人类 镜片和经过各种物理化学处理的镜片 压力 这些研究很好地使我们能够发展一个深入的 膜性能相对细微变化的评价 这会导致膜性能的显著变化 有助于白内障的形成。 这项研究的结果将 加强必要的知识基础，以确保未来 人类白内障药物治疗的进展。 这项研究的具体目标如下： 1. 测量初级红外和拉曼光谱 从正常人中获得的脂囊泡分散体的特征 牛和人的晶状体。 光谱特征将 包括：a）流动性和相变温度; B） 正交或单斜堆积顺序; c）脂头部基团 相互作用; d）脂酰基界面区有序和堆积; e）定量脂质饱和度; f）确认脂质组成。 囊泡分散体的膜脂质组成将是 薄层色谱法和高效液相色谱法测定 液相色谱法 2. 确定内外膜的影响 蛋白质对透镜细胞光谱特性的影响 膜。 3. 探索光谱特征的变化， 透镜的老化和人类白内障。 4. 确定氧化应激、紫外线暴露， 钙和胆固醇结合抗生素菲律宾对 透镜细胞膜和膜的光谱特性 脂质。 5. 膜脂光谱相关性探讨 具有透镜膜性能的特性，即ATP酶 活性和膜泡溶质通量。

The overall objective of our proposed investigation is to use state-of-the-art spectroscopic techniques to examine regional conformational; compositional and structural characteristics of lens membrane lipids and proteins. Spectroscopic findings could be related to membrane composition and performance. We will investigate normal bovine and human lenses, cataractous human lenses and lenses subjected to a variety of physicochemical stresses. Such studies well permit us to develop an in depth appreciation of relatively subtle changes in membrane properties which can lead to considerable changes in membrane performance contributing to cataractogenesis. The results of this study will strengthen the foundation of knowledge necessary to assure future development of medical therapy of human cataract. The specific aims of this study are as follows: 1. Measure primary infrared and Raman spectroscopic characteristics of lipid vesicle dispersions obtained from normal bovine and human lenses. Spectroscopic characteristics will include: a) fluidity and phase transition temperatures; b) orthorhombic or monoclinic packing order; c) lipid head group interactions; d) lipid acyl-interface region order and packing; e) quantitate lipid saturation; f) confirm lipid composition. The membrane lipid composition of the vesicle dispersions will be determine by thin layer chromatography and high performance liquid chromatography. 2. Determine the influence of intrinsic and extrinsic membrane proteins upon spectroscopic characteristics of lens cell membranes. 3. Explore changes in spectroscopic characteristics accompanying aging of the lens and in human cataract. 4. Determine the impact of oxidative stress, UV light exposure, calcium and the cholesterol binding antibiotic filipin upon the spectroscopic characteristics of lens cell membranes and membrane lipids. 5. Explore the correlation of membrane lipid spectroscopic characteristics with lens membrane performance i.e. ATPase activity and membrane vesicle solute flux.