MICRODISSECTED CATARACTOUS LENSES

描述（申请人的摘要）：本发明的长期目标 建议是更好地了解生物化学和生物物理变化 随着年龄的增长，人类透镜中发生的变化，最终导致 白内障 拟议的研究将有助于实现， 未来，最终目标是大大延缓形成 老年性白内障以及消除继发性白内障的形成 白内障是白内障手术后常见的疾病。 具体目标I：表征人透镜上皮蛋白：一个关键 在维持透镜体内平衡中起作用的是上皮层。 白内障后获得的新鲜切除的人上皮 手术将被使用。 将对个体进行测量 上皮 一部分上皮将用于酶活性 而剩下的部分将用于确定 的mRNA水平的特定酶使用PCR技术。 的 人上皮细胞cDNA文库的构建将使我们能够表征 各种人类上皮蛋白，并找出是否有透镜特异性 任何一种蛋白质的异构体 具体目标II：阐明新发现的分子伴侣 α-晶体蛋白的功能：分子伴侣在 抑制变性蛋白质的聚集，这是老化透镜的性质 需求 我们发现α-晶状体蛋白是一种分子伴侣。 这 表明它在透镜中具有双重作用。 一个作为折射元件 第二个作为伴侣蛋白，保护其他蛋白质免于聚集。 α-晶体蛋白的物理性质将通过各种方法进行研究。 技术. α-晶状体蛋白与其他透镜蛋白的相互作用 如beta，gamma和MP-26等。 衰老的影响， 白内障对α-晶状体蛋白伴侣功能的影响将在 正常和白内障的人类晶状体。

DESCRIPTION (Applicant's abstract): The long term objective of this proposal is to better understand the biochemical and biophysical changes that occur in the human lens with aging, and which ultimately lead to cataract. The proposed studies will be useful for achieving, in the future, the ultimate goal of significantly retarding the formation of aging-dependent cataract as well as eliminating the formation of secondary cataract which is commonly encountered following cataract surgery. Specific Aim I: To characterize human lens eptithelial proteins: A key player in the maintenance of lens homeostasis is the epithelial layer. Freshly excised human epithelium which will be obtained following cataract surgery will be used. Measurements will be performed on individual epithelium. A portion of the epithelium will be use for enzyme activity determination, whereas the remaining piece will be used for determination of mRNA levels of the specific enzyme using PCR technology. The construction of human epithelial cDNA library will allow us to characterize various human epithelial proteins, and to find if there are lens- specific isoforms for any of the proteins. Specific Aim II: To elucidate the newly discovered molecular chaperone function of alpha-crystallins: Molecular chaperones are very efficient in suppressing aggregation of denatured protein, a property the aging lens needs. Alpha-crystallin was found by us to be a molecular chaperone. This suggests that it has a dual role in the lens. One as a refractive element and a second one as a chaperone protecting other proteins from aggregation. The physical properties of alpha-crystallins will be studied by various techniques. The interaction of alpha-crystallin with other lens proteins such as beta, gamma, and MP-26 will be evaluated. The effects of aging and cataract on the chaperone function of alpha-crystallin will be examined in normal and cataractous human lenses.