MASS SPECTROMETRIC ANALYSIS OF LENS PROTEINS

描述：晶体蛋白是透镜的主要蛋白质， 在透镜成熟和老化期间的部分分裂。 晶体蛋白裂解 都发生在动物和人类的镜头里。 因为动物的白内障 这一进程的加速，主要是 研究人员推测，人类白内障也可能是由于 晶状体蛋白的不适当切割。 为了验证这一假设，PI 提出：1）分离和表征未鉴定的蛋白水解酶 负责人透镜中大部分蛋白质片段化的酶。 2)在二维电泳凝胶上产生详细的图谱 在动物中发现的完整和修饰的透镜蛋白的同一性， 人类晶状体 3)创建标准化的二维电泳 透镜蛋白数据库，可在万维网上访问。这项工作将 使用基质辅助激光解吸质谱进行 蛋白酶底物分析和二维电泳 分离的透镜蛋白。 这些研究很重要，因为1）酶 导致人类透镜晶体蛋白破碎的原因尚不清楚; 2）很少 比较动物蛋白质的身份和修饰的信息 和人类晶状体现在存在;和3）没有机制，易于共享透镜 研究人员之间的蛋白质数据可用。 这一长期目标 项目是：改进用于研究蛋白质修饰的方法， 透镜，开发适当的动物模型来研究人类白内障，测试是否 蛋白质碎片化可能是人类白内障的一个原因，并产生蛋白酶 抑制剂作为可能的抗白内障药物。

DESCRIPTION: Crystallins, the major proteins of the lens, undergo rapid partial cleavage during lens maturation and aging. Cleavage of crystallins takes place in both animal an human lenses. Since cataracts in animals have been associated with an acceleration of this process, the principal investigator hypothesizes that human cataracts may also result from the inappropriate cleavage of crystallins. To test this hypothesis the PI proposes: 1) Isolating and characterizing the unidentified proteolytic enzyme responsible for the majority of protein fragmentation in human lens. 2) Producing detailed maps on two-dimensional electrophoresis gels of the identities of both intact and modified lens proteins found in animal and human lenses. 3) Creating a standardized two-dimensional electrophoresis database for lens proteins for access on the World Wide Web. This work will be performed using matrix assisted laser desorption mass spectrometric analysis of protease substrates and two-dimensional electrophoretically separated lens proteins. These studies are important because 1) the enzyme causing the fragmentation of human lens crystallins is unknown; 2) little information comparing the identities and modifications of proteins in animal and human lenses now exists; and 3) no mechanism for easily sharing lens protein data between researchers is available. The long term goals of this project are to: improve methods used to study protein modifications in the lens, develop appropriate animal models to study human cataract, test if protein fragmentation may be a cause of human cataract, and develop protease inhibitors as possible anticataract drugs.