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-钙离子依赖性二酰基甘油调节蛋白 磷酸化（PKC），以及 磷脂酰肌醇、磷脂酰肌醇-4-磷酸和 将在家兔中研究磷脂酰肌醇-4，5-二磷酸 角膜测试假设，肌醇脂质循环和 蛋白质磷酸化反应是相互关联的， 角膜中的受体激活阶段，并参与作为 角膜创伤过程中细胞内生长调控途径 治愈 此外，生长因子可能调节角膜上皮细胞的生长， 通过控制肌醇脂质循环和蛋白质 角膜中的磷酸化。 组蛋白H1将被用作 外源底物以测定蛋白激酶C。 此外，内源性 磷酸化，包括酪氨酸磷酸化， 通过评估创伤后的不同时间段， 钙-磷脂敏感性和肿瘤促进剂-磷脂 活性在可溶性和颗粒部分。 这种设计 将使我们能够跟踪可溶性和颗粒的时间进程 创伤愈合过程中蛋白质磷酸化， 这些事件与肌醇脂质循环有关。 两款车型将 使用的是一种机械模型，其中上皮细胞被选择性地 受伤的，和一个低温模型，其中的三层， 角膜受到影响。 在这两种模型中，蛋白质和脂质 磷酸化将在细胞的过程中进行， 上皮细胞、间质细胞和 内皮细胞 第一个模型可以让我们研究生物化学 当上皮受伤时其他层的改变， 以及这些改变影响愈合的程度 上皮 第二个模型可以让我们研究 间质和内皮以及上皮细胞。 与细胞运动和增殖的相关性将通过 组织学、DNA和伤口半径测量。 高性能液体等生化技术 色谱法、毛细管气液色谱法和低- 并将使用高压电泳。 的结果 建议将提供更好的了解生物化学 肌醇脂质和蛋白质磷酸化在角膜和 将有助于角膜伤口愈合的管理-通过 产生关于细胞新的受体后通路的信息 信号传导，其中药物可能具有调节 角膜细胞中细胞间通讯的破坏 伤人