CATARACT RELATED MODIFICATION OF LENS CRYSTALLINS

描述(研究人员摘要)：白内障，一种常见的疾病 老龄化，在患有尿毒症的人中发病率增加， 糖尿病，或慢性腹泻。预防犯罪的理性发展 建议和治疗药物一直受到缺乏 关于晶状体共价修饰的分子水平知识 导致或伴随白内障的蛋白质。建议数 调查将通过使用新的分析方法来满足这一需求 用质谱学方法分析正常、老年人和老年人的蛋白质 患有白内障的人类晶状体。在第一层次的分析中，一种新的形式 电喷雾电离质谱仪(ESIMS)，将 用于测定晶状体蛋白质的相对分子质量 0.02%的精度(5个质量单位)。然后晶状体蛋白质将被 蛋白质分解成的多肽，其相对分子质量将是 用直接耦合微孔测定，测量精度为0.3质量单位 高效液相色谱快原子轰击质谱仪。其相对分子质量 晶状体蛋白和蛋白水解肽及其层析 和电泳学性质，将被用来鉴定，在分子 水平、正常和共价修饰的晶状体蛋白。对一个池的结果 正常的人类晶状体将被用来建立一个数据库，结果是 从类似的分析中，我们将对老年性和白内障晶状体进行比较。 参考文献中未包含分子量的蛋白质和多肽 数据库将被视为已进行共价修改。 与尿毒症、糖尿病和慢性腹泻相关的白内障晶状体 将通过相同的程序进行研究，这样共价修饰的蛋白质 每种类型的白内障都可以识别出独特的多肽。镜头 蛋白质经历了独特的老化和共价修饰 具体形式的白内障将通过各种微观手段进行调查 在分子水平上鉴定细菌类型的分析方法 以及蛋白质上具有 进行了修改。

DESCRIPTION (Investigator's Abstract): Cataract, a common disorder of aging, occurs with increased incidence among persons afflicted with uremia, diabetes, or chronic diarrhea. The rational development of preventive recommendations and therapeutic drugs has been hampered by a lack of knowledge at the molecular level about covalent modifications to lens proteins that either cause or accompany cataract. The proposed investigation will address this need by using new analytical methods based on mass spectrometry to analyze proteins isolated from normal, aged, and cataractous human lenses. In the first level of analysis, a new form of mass spectrometry, electrospray ionization mass spectrometry (ESIMS), will be used to determine the molecular weights of lens proteins with an accuracy of 0.02 percent (5 mass units). Lens proteins will then be proteolytically fragmented into peptides whose molecular weights will be determined with an accuracy of 0.3 mass units by directly-coupled microbore HPLC fast atom bombardment mass spectrometry. The molecular weights of the lens proteins and proteolytic peptides, as well as their chromatographic and electrophoretic properties, will be used to identify, at the molecular level, normal and covalently modified lens proteins. Results for a pool of normal human lenses will be used to make a data bank with which results from similar analyses of aged and cataractous lenses will be compared. Proteins and peptides with molecular weights that are not in the reference data bank will be considered to have undergone covalent modification. Cataractous lenses associated with uremia, diabetes, and chronic diarrhea will be studied by the same procedure so that covalently modified proteins and peptides unique to each type of cataract can be discerned. Lens proteins that have undergone covalent modifications unique to aging and specific forms of cataract will be investigated by a variety of micro analytical methods to identify at the molecular level the type of modification as well as the specific site on the protein which has undergone modification.