ROLE OF PROTEIN AND LIPID PHOSPHORYLATION IN CORNEA

磷脂-钙依赖的二酰甘油调节蛋白 磷酸化(PKC)，以及蛋白的代谢 磷脂酰肌醇、磷脂酰肌醇-4-磷酸和 磷脂酰肌醇-4，5-二磷酸将在兔体内进行研究 以检验肌醇脂质循环和眼角膜 蛋白质的磷酸化反应是相互关联的 在角膜中处于受体激活阶段，并参与了 角膜损伤过程中的细胞内生长调控途径 治愈。此外，可能是生长因子调节了角膜。 通过控制肌醇脂质循环和蛋白质促进伤口愈合 角膜中的磷酸化。组蛋白H1将被用作 外源性底物测定蛋白激酶C 磷酸化，包括酪氨酸磷酸化，将是 通过评估伤后不同时间段进行评估 钙磷脂敏感性与肿瘤促进剂磷脂 可溶组分和颗粒组分中的活性。这个设计 将使我们能够跟踪可溶性和颗粒物的时间进程 创伤愈合过程中蛋白质的磷酸化及其相互关系 这些事件与肌醇脂质循环有关。将有两款车型 使用的是一种机械模型，在该模型中，上皮细胞被选择性地 和一个低温模型，在这个模型中，三层 角膜受到影响。在这两个模型中，蛋白质和脂肪 在细胞的过程中，会伴随着磷酸化。 上皮、间质和 内皮细胞。第一个模型将使我们能够研究生化 当上皮损伤时，其他层的变化， 以及这些改变对伤口愈合的影响程度 上皮组织。第二个模型将使我们能够研究治疗 间质和内皮细胞以及上皮细胞。 与细胞运动和增殖的相关性将通过 组织学、DNA和创面半径测量。 高效液体等生化技术 柱层析、毛细管气相液相色谱、低... 并将使用高压电泳法。这样做的结果 提案将提供对生物化学的更好的理解 角膜中的肌醇脂类和蛋白质磷酸化 将在角膜伤口愈合的管理中有用。 产生关于细胞新的受体后通路的信息 信号，其中药物可能有可能调节 术后角膜细胞间通讯中断 伤人。

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.