ROLE OF PROTEIN AND LIPID PHOSPHORYLATION IN CORNEA

角膜中蛋白质和脂质磷酸化的作用

• 批准号: 3263106
• 负责人: Haydee E.P. Bazan
• 金额: $ 12.14万
• 依托单位: LSU HEALTH SCIENCES CENTER
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-09-30 至 1992-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3263106
• 关键词: autoradiography; biochemistry; biological signal transduction; calcium; cell differentiation; cell growth regulation; cell motility; chromatography; cornea disorder; corneal endothelium; corneal epithelium; corneal stroma; diacylglycerols; electrophoresis; epidermal growth factor; eye injury; growth factor; histology; inositol; laboratory rabbit; lipid metabolism; peptidases; phorbols; phosphatidylinositols; phosphorylation; protein kinase C; radioassay; tumor promoters; tyrosine; wound healing

Phospholipid-Ca2+ dependent, diacylglycerol modulated protein phosphorylation (PKC), as well as the metabolism of phosphatidylinositol, phosphatidylinositol-4-phosphate and phosphatidylinositol-4,5-bisphosphate, will be studied in the rabbit cornea to test the hypothesis that the inositol lipid cycle and protein phosphorylation reactions are interrelated at the post- receptor activation stage in the cornea, and are involved as intracellular growth control pathways during corneal wound healing. Also, it may be that growth factors modulate corneal wound healing by controlling the inositol lipid cycle and protein phosphorylation in the cornea. Histone H1 will be used as an exogenous substrate to assay protein kinase C. Also, endogenous phosphorylation, including tyrosine phosphorylation, will be evaluated for various time periods after wounding by assessing calcium-phospholipid sensitivity and tumor promoter-phospholipid activity in both the soluble and particulate fractions. This design will enable us to follow the time course of soluble and particulate protein phosphorylation during wound healing and to correlate these events with the inositol lipid cycle. Two models will be used, a mechanical model in which the epithelium is selectively wounded, and a cryogenic model in which the three layers of the cornea are affected. In both models, protein and lipid phosphorylation will be followed during the process of cell migration and profileration in the epithelium, stroma and endothelium. The first model will allow us to study biochemical alterations in the other layers when the epithelium is wounded, and the extent to which these alterations affect the healing of the epithelium. The second model will allow us to study the healing of the stroma and endothelium as well as the epithelium. Correlation with cell movement and proliferation will be made by histology, DNA and radius of the wound measurement. Biochemical techniques such as high performance liquid chromatography, capillary gas liquid chromatography, and low- and high-voltage electrophoresis will be used. The results of this proposal will provide a better understanding of the biochemistry of inositol lipids and protein phosphorylation in the cornea and will be useful in the management of corneal wound healing- by generating information on new postreceptor pathways of cell signaling, where drugs may have the potential to modulate the breakdown of cell-to-cell communication in cornea cells after wounding.

磷脂-CA2+依赖性二酰基甘油调制蛋白 磷酸化（PKC）以及代谢 磷脂酰肌醇，磷脂酰肌醇-4-磷酸和 磷脂酰肌醇-4,5-双磷酸盐将在兔子中进行研究 角膜以检验肌醇脂质周期和 蛋白质磷酸化反应在后期相互关联 角膜中的受体激活阶段，并参与 角膜伤口期间细胞内生长控制途径 康复。 而且，可能是生长因子调节角膜 通过控制肌醇脂质周期和蛋白质来愈合伤口 角膜中的磷酸化。 组蛋白H1将用作 外源性底物进行测定蛋白激酶C.，内源性 磷酸化，包括酪氨酸磷酸化，将是 通过评估在伤害后评估各个时间段 钙磷脂敏感性和肿瘤启动子磷脂 可溶性和颗粒分数的活性。 这个设计 将使我们能够遵循可溶性和颗粒物的时间过程 伤口愈合过程中的蛋白质磷酸化并相关 这些事件具有肌醇脂质周期。 两个模型将是 使用的是一种机械模型，上皮有选择性 受伤和一个低温模型，其中三层 角膜受到影响。 在这两个模型中，蛋白质和脂质 在细胞过程中将遵循磷酸化 上皮，基质和 内皮。 第一个模型将使我们能够研究生化 上皮受伤时其他层的改变， 以及这些改变影响康复的程度 上皮。 第二个模型将使我们能够研究康复 基质和内皮以及上皮。 与细胞运动和增殖的相关性将由 伤口测量的组织学，DNA和半径。 高性能液体等生化技术 色谱，毛细管气体色谱和低 - 将使用高压电泳。 结果的结果 建议将更好地了解生物化学 肌醇脂质和角膜中的蛋白质磷酸化 通过 生成有关细胞的新后训练途径的信息 信号传导，药物可能具有调节的潜力 在角膜细胞中细胞对细胞通信的崩溃之后 受伤。