SPECTROSCOPIC & RELATED STUDIES ON LENS MEMBRANE LIPIDS

本建议的总体目标是研究区域 透镜膜的形态、组成和结构特征 脂质和蛋白质，并将膜组成与膜结构联系起来 和性能（即渗透性、阳离子泵活性）。 我们将使用 表征正常家兔的最新光谱技术 和人类透镜膜，并将它们与白内障人类透镜进行比较 膜和经受各种生理化学应力的膜。 我们的假设是，膜脂组成的改变， 结构影响膜性能。 拉曼和红外 光谱学是将用于实现六个目标的工具之一， 来验证我们的假设 组织：将针对晶状体区域研究人类晶状体 随年龄和白内障类型而变化。 L.以上组织中的脂质头基成分是什么？ 是未知脂质的一级结构，其占脂质的60%。 磷脂？ 2.上述组织中的脂质氧化程度如何？ 氧化 将通过测量：脂质-OH和-OOH，胆固醇 修饰、绿色和蓝色荧光团以及饱和度。 3.内源性和外源性蛋白质是如何影响血脂和 脂质-蛋白质相互作用， 上述组织中膜蛋白的结构差异？ 4.什么是影响：氧化应激，钙和 胆固醇/菲律宾脂对透镜池光谱特性的影响 膜和膜脂？ 5.脂质氧化的次级产物是否会改变次级和 纯化的牛晶状体蛋白或人透镜的三级结构 匀浆？ 6.膜脂组成与结构的相关性是什么 具有透镜膜性能，即Ca-ATPase、Na，K-ATPase和MIP 26 功能？

The overall objective of this proposal is to study the regional conformation, composition and structural characteristics of lens membrane lipids and proteins, and relate membrane composition to membrane structure and performance (i.e. permeability, cation pump activity). We will use state of the art spectroscopic techniques to characterize normal rabbit and human lens membranes and to compare them to cataractous human lens membranes and membranes subjected to a variety of physiochemical stresses. Our hypothesis is that alterations in membrane lipid composition and structure influence membrane performance. Raman and infrared spectroscopies are among the tools that will be used to pursue six aims to test our single hypothesis. Tissues: Human lenses will be studied with respect to lens-regional changes with age and cataract type. l. What is the lipid headgroup composition in the tissues above and what is the primary structure of the unknown lipid that comprises 60% of the phospholipids? 2. What is the degree of lipid oxidation in the tissues above? oxidation will be assessed by measuring: lipid -OH and -OOH, cholesterol modifications, green and blue fluorophores and saturation. 3. How do intrinsic and extrinsic proteins influence lipid-lipid and lipid-protein interactions and, are there secondary and tertiary structural differences in membrane proteins in the tissues above? 4. What is the impact of: oxidative stress, calcium and cholesterol/filipin upon the spectroscopic characteristics of lens cell membrane and membrane lipids? 5. Do secondary products of lipid oxidation alter the secondary and tertiary structure of purified bovine crystallins or human lens homogenates? 6. What is the correlation of membrane lipid composition and structure with lens membrane performance, i.e Ca-ATPase, Na, K-ATPase and MIP26 function?