REGULATION OF PROTEIN PHOSPHORYLATION IN THE EYE

眼睛中蛋白质磷酸化的调节

• 批准号: 3259819
• 负责人: JAMES A NATHANSON
• 金额: $ 10.16万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3259819
• 关键词: adenylate cyclase; beta adrenergic agent; calcium; chromatography; cyclic AMP; cyclic GMP; cyclic nucleoside monophosphate; enzyme substrate; gel electrophoresis; guanylate cyclase; histochemistry /cytochemistry; homeostasis; hormone regulation /control mechanism; immunochemistry; lens; lens proteins; phosphorylation; prostaglandins; protein kinase; scintillation counter; uvea ciliary body

Phosphorylation of intracellular proteins is a key biochemial mechanism by which many extracellular hormones exert their effects on cellular function. In many instances, calcium ions or cyclic nucleotides (cyclic AMP and cyclic GMP) act as intracellular intermediaries between hormone binding and activation of phosphorylation. Surprisingly, although much work has been done on protein phosphorylation mechanisms in the retina, little is known about the regulation of phosphorylation in the ciliary process and lens, including its epithelium. Hormone receptors (beta-adrenergic, prostaglandin, steroid, parathyroid hormone) are present in these two tissues, and preliminary work (including data from this laboratory) indicates the existence of endogenous protein substrates that are phosphorylated by cyclic-nucleotide-dependent protein kinases. We propose to investigate, in the lens, lens capsule and ciliary process, the regulation of protein phosphorylation by cyclic nucleotides, calcium and various hormones. Using both intact tissue and broken cell preparations, endogenous protein substrates for phosphorylation will be identified and characterized, along with the phosphorylating (kinases) and dephosphorylating (phosphatases) enzymes. In the ciliary process, phosphorylation in the epithelium, itself, will be studied using epithelial cell isolation techniques. Particular emphasis will be put on the hormone regulation of phosphorylation using prelabeling and back-phosphorylation techniques. These studies should provide an improved understanding of how hormones and ions regulate these tissues. In the ciliary process, such regulation is relevant to the physiology of aqueous humor secretion (with relevance to glaucoma) and, in the lens epitheliun, to the maintenance of lens metabolism (with relevance to cataract formation).

细胞内蛋白的磷酸化是一种关键的生化机制 许多细胞外激素对细胞的影响 功能。 在许多情况下，钙离子或环状核苷酸（环状 AMP和环状GMP）充当激素之间的细胞内中介 磷酸化的结合和激活。 令人惊讶的是，虽然很多 已经完成了视网膜中蛋白质磷酸化机制的工作， 关于纤毛中磷酸化的调节知之甚少 过程和镜头，包括其上皮。 激素受体 （β-肾上腺素能，前列腺素，类固醇，甲状旁腺激素）存在 在这两个组织和初步工作中（包括来自此的数据 实验室）表明存在内源性蛋白底物 被环核苷酸依赖性蛋白激酶磷酸化。 我们建议在镜头，镜头胶囊和睫状过程中调查 循环核苷酸，钙调节蛋白质磷酸化 和各种激素。 使用完整的组织和破裂的细胞 制剂，用于磷酸化的内源性蛋白质底物将是 鉴定和表征，以及磷酸化（激酶）和 去磷酸化（磷酸酶）酶。 在睫状过程中， 上皮本身的磷酸化将使用上皮研究 细胞隔离技术。 特别重点将放在激素上 使用前标签和后​​磷酸化调节磷酸化 技术。 这些研究应提高人们对如何 激素和离子调节这些组织。 在睫状过程中 调节与水性幽默分泌的生理学有关（与 与青光眼相关），在镜头上皮中与维持 晶状体代谢（与白内障形成有关）。