ASCORBIC ACID GLYCATION AND SENILE CATARACT FORMATION

抗坏血酸糖化与老年性白内障形成

• 批准号: 6384577
• 负责人: BERYL J ORTWERTH
• 金额: $ 27.38万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6384577
• 关键词: SDS polyacrylamide gel electrophoresis; aging; animal tissue; ascorbate; binding proteins; cataract; crosslink; disease /disorder prevention /control; enzyme linked immunosorbent assay; free radical oxygen; glycation; high performance liquid chromatography; human tissue; ionophores; lens proteins; mass spectrometry; molecular pathology; nuclear magnetic resonance spectroscopy; organ culture; pathologic process; protein protein interaction; protein structure; thin layer chromatography

In the last two decades, major advances have been made in understanding lens protein structure and lens crystallin gene expression. We are obtaining increasingly detailed knowledge about cataracts in mice and rats, but the mechanisms that result in age-onset cataract in humans, still remains elusive. Many in vitro experiments mimic features seen in these cataracts, but there is little in the way of confirmation that these reactions are significant, or causal in the process. Our laboratory has been investigating the role of advanced glycation endproducts in this process. Specifically, we are investigating whether the oxidation products of ascorbic acid are responsible for the protein-protein crosslinks, chromophores and fluorophores seen in these lenses. In the work described here we hope to find common structures between calf lens proteins ascorbylated in vitro and the protein modifications seen in aged human lenses and cataracts obtained from India. A scheme for the isolation of these modifications from proteins is presented. Also, the common compounds will be identified by mass spectrometry, their spectral properties and proton NMR. In addition, we intend to study the degradation of ascorbic acid in cultured human lenses, and to measure the effect of external agents to bring about the oxidation and incorporation of ascorbic acid into the proteins in these lenses.

在过去的二十年中，在理解透镜蛋白结构和透镜晶体蛋白基因表达方面取得了重大进展。 我们正在获得关于小鼠和大鼠白内障的越来越详细的知识，但导致人类老年性白内障的机制仍然难以捉摸。 许多体外实验模拟了这些白内障中观察到的特征，但几乎没有证据表明这些反应是显着的，或者在这个过程中是因果关系。 我们的实验室一直在研究晚期糖基化终产物在这一过程中的作用。 具体来说，我们正在研究是否抗坏血酸的氧化产物是负责蛋白质-蛋白质交联，发色团和荧光团在这些镜头中看到。 在这里描述的工作中，我们希望找到体外抗坏血酸化的小牛透镜蛋白质与从印度获得的老年人晶状体和白内障中看到的蛋白质修饰之间的共同结构。 提出了从蛋白质中分离这些修饰的方案。 此外，常见的化合物将通过质谱法，其光谱特性和质子NMR进行鉴定。 此外，我们打算研究抗坏血酸在培养的人晶状体中的降解，并测量外部试剂的作用，以使抗坏血酸氧化并掺入这些晶状体中的蛋白质中。