ATP-LINKED EFFECTORS OF NA,K-ATPASE AND CATARACT

NA,K-ATP酶和白内障的ATP连锁效应子

• 批准号: 3263814
• 负责人: MARGARET H GARNER
• 金额: $ 19.57万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-07-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3263814
• 关键词: adenosine diphosphate; adenosine triphosphate; adenosinetriphosphatase; aging; amino group; cataract; conformation; cow; diabetic cataract; diabetic ophthalmopathy; glucose; glycosylation; high performance liquid chromatography; human tissue; isozymes; laboratory rabbit; laboratory rat; lens; membrane permeability; molecular site; nuclear magnetic resonance spectroscopy; organ culture; ouabain; oxidation; phosphates; radioassay; sodium potassium exchanging ATPase; uremias

Changes in lenticular Na+ and K+ concentrations have been correlated with increasing lens color and increasing cortical opacity. Na,K-ATPase is inhibited in epithelium in 37% of the cataract population. Inhibition of the Na,K-ATPase of the lens cortex is more general, occurring in 80-90 percent of the lenses. The purpose of the proposed studies is to characterize the Na,K- ATPase of the epithelium and cortex of normal and cataractous lenses. Since human cataract lenses are not available for study, animal models and in vitro model systems will be employed. During the next five years, the sites of glucosylation, carbamylation, and oxidation of lens epithelial cell Na,K-ATPase will be located. The interaction of Na+,K+,Mg2+,ATP,ADP,Pi, and ouabain with the three modified Na,K-ATPase preparations will be determined. This will be accomplished using steady state kinetics techniques, lens organ culture and lens epithelial cell culture techniques as well as non steady state kinetics techniques with purified renal Na,K-ATPase in synthetic membrane vesicles. Studies will be initiated to characterize the Na,K-ATPase of the superficial cortical fibers. The isozyme(s) present in the cortex will be determined using specific antisera to the three major sequenced heavy subunit isozymes. The fate of the Na,K-ATPase in the lens cortex of selected model cataracts will be determined.

晶状体Na+和K+浓度的变化 与增加镜片颜色和增加皮质层相关 不透明。37%的人上皮细胞中Na，K-ATPase受到抑制 白内障人口。晶状体Na，K-ATPase的抑制作用 皮质更普遍，出现在80%-90%的晶状体中。 拟议研究的目的是表征Na，K- 正常和白内障的上皮和皮质中的ATPase 隐形眼镜。由于人类白内障晶状体不能用于研究， 将采用动物模型和体外模型系统。 在接下来的五年里，糖基化的部位， 晶状体上皮细胞Na，K-ATPase的氨甲基化和氧化 将会被找到。Na~+、K~+、Mg~(2+)、ATP、ADP、PI、 和哇巴因与三种修饰的Na，K-ATPase制剂 将会被确定。这将使用稳定状态来完成 动力学技术、晶状体器官培养和晶状体上皮细胞 培养技术和非稳态动力学技术 在合成膜囊泡中加入纯化的肾脏Na，K-ATPase。 将启动研究，以确定Na，K-ATPase的特征 浅层皮质纤维。皮层中存在的同工酶(S) 将用三种主要病毒的特异性抗血清来确定 对重亚基同工酶进行了测序。Na，K-ATPase的命运 在选定的模型白内障的晶状体皮质中将进行测定。