PHYSICAL AND CHEMICAL BASIS OF LENS OPACITY

晶状体混浊的物理和化学基础

• 批准号: 7084495
• 负责人: GEORGE B BENEDEK
• 金额: $ 39.06万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-05-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7084495
• 关键词: anterior chamber; atomic force microscopy; bioenergetics; cataract; cell osmotic pressure; chemical aggregate; clinical research; covalent bond; crystallins; gel electrophoresis; intraocular fluid; ionic strengths; lens; lens proteins; light microscopy; light scattering; mathematical model; molecular pathology; molecular weight; phase change; protein protein interaction; site directed mutagenesis; spectrometry; thermodynamics

Our long-term objective is to identify the molecular factors responsible for light scattering and opacification of the lens. Light scattering can be produced in the lens by the formation of protein aggregates, the condensation of proteins into coexisting protein-rich and protein-poor liquid phases and the condensation of proteins into crystalline solid phases. In previous years, we have identified various molecular mechanisms that produce such condensates, and thereby result in light scattering and opacification in gamma crystallin solutions in vitro. Some of these mechanisms are now known to be responsible for several human genetic cataracts. We now wish to extend our work by conducting investigations of the condensates formed by the beta crystallins. Moreover, with the extensive knowledge base of gamma crystallins, we also want to explore the thermodynamic and kinetic behavior of the mixtures of beta and gamma crystallins, in an effort to emulate more realistically the situation present in the lens cytoplasm. To achieve these objectives, we propose the following Specific Aims: 1. To determine the phase diagrams, and the kinetics of aggregation of aqueous solutions of recombinant human beta crystallins, and their naturally occurring mutant and truncated forms. 2. To explore the interactions between beta and gamma crystallins, by studying the phase-diagrams and kinetics of aggregation of solutions of beta and gamma crystallin mixtures. 3. To develop a theoretical description which can predict the phase diagrams, and the kinetics of aggregation for solutions of pure beta and gamma crystallins as well as their mixtures.

我们的长期目标是确定负责镜头光散射和不透明的分子因素。可以通过形成蛋白质聚集体，将蛋白质凝结成富含蛋白质的蛋白质和蛋白质贫乏的液相，以及将蛋白质凝结成结晶固体相的凝结。在过去的几年中，我们已经确定了产生这种冷凝水的各种分子机制，从而导致体外伽马晶体溶液中的光散射和无情。现在已知其中一些机制负责几种人类遗传性白内障。现在，我们希望通过对由该工作进行调查来扩展我们的工作 β结晶蛋白。此外，借助伽马晶蛋白的广泛知识基础，我们还希望探索β和伽马晶蛋白混合物的热力学和动力学行为，以便更真实地模仿镜片细胞质中的情况。为了实现这些目标，我们提出以下特定目的： 1。确定重组人β结晶蛋白水溶液的聚集的相图以及它们天然存在的突变体和截断形式。 2。通过研究β和γ晶体混合物溶液的聚集的相结合和动力学来探索β和伽马晶体之间的相互作用。 3。开发一个可以预测相图的理论描述，以及用于纯β和伽马晶体溶液及其混合物的溶液的聚集动力学。