REGULATION OF CALMODULIN DEPENDENT PROTEIN KINASES

钙调蛋白依赖性蛋白激酶的调节

• 批准号: 2037262
• 负责人: ARTHUR M EDELMAN
• 金额: $ 21.36万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-02-01 至 1999-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2037262
• 关键词: PC12 cells; biological signal transduction; calcium flux; calmodulin dependent protein kinase; chemical kinetics; enzyme activity; enzyme induction /repression; immunocytochemistry; neuropharmacologic agent; neuropharmacology; phosphoproteins; phosphorylation

DESCRIPTION: Extracellular signal-control elevations in intracellular Ca2+ levels regulate a diverse array of cellular phenomena. Evidence has accumulated for calcium dyshomeostatsis in nervous system pathologies including HIV-induced neuronal injury, Alzheimer's disease and Parkinson's disease suggesting the potential for new pharmacological interventions for these disorders based on alteration in intracellular Ca2+ levels. Calcium calmodulin (CaM) dependent protein kinases (CaM kinase) are important mediators of the effects of elevated levels of intracellular calcium upon physiologically relevant target proteins. CaM kinase I is the focus of this proposal. This research is intended to provide an integrated approach for elucidating the signaling cascade for phosphorylation and activation of CaM kinase I. The specific aims are: 1. Identification of extracellular and intracellular signals which induce phosphorylation and activation of CaM kinase I in pheochromocytoma PC12 cells; 2, Analysis of signal transduction mechanisms in the CaM kinase I cascade. Specifically to determine by immunoprecipitation techniques if CaM kinase I (or CaM kinase IV) forms complexes with its activating kinases (CaMKIKs) and evaluate whether phosphorylation of CaMKIKs influence their abilities to phosphorylate and activate CaM kinase I and to form complexes with CaM kinase I.; 3. Evaluation of the respective roles of CaMKIKs in CaM kinase cascades. Specifically to determine through studies of their respective kinetics of phosphorylation whether CaMKIKs utilize CaM kinase I preferentially as substrate relative to CaM kinase IV and determine their respective localization using Northern and immunoblotting and immunocytochemistry, and whether the CaMkKIKs have cell type or tissue specific functions.

描述：细胞外CA2+的细胞外信号控制高程 水平调节各种细胞现象。 证据有 在神经系统病理中积累了钙dyshomeostatsis 包括艾滋病毒引起的神经元损伤，阿尔茨海默氏病和帕金森氏病 疾病表明有可能进行新的药理学干预措施 这些疾病基于细胞内Ca2+水平的改变。 钙钙调蛋白（CAM）依赖性蛋白激酶（CAM激酶）为 重要的介体的细胞内水平升高的影响 生理相关靶蛋白的钙。 凸轮激酶我是 该提议的重点。 这项研究旨在提供综合的 阐明信号级联的方法，以进行磷酸化和 CAM激酶I的激活I。 具体目的是：1。鉴定细胞外和 诱导CAM磷酸化和激活的细胞内信号 嗜铬细胞瘤PC12细胞中的激酶I； 2，信号转导的分析 CAM激酶I级联的机制。 专门确定 免疫沉淀技术如果CAM激酶I（或CAM激酶IV）形成 与其激活激酶（CAMKIK）的复合物，并评估是否 Camkiks的磷酸化影响了它们磷酸化和 激活CAM激酶I并用CAM激酶I。 3。 评估CAMKIK在CAM激酶级联反应中的各自作用。 特别是通过研究其各自的动力学的研究 Camkiks是否优先使用CAM激酶I，因为 相对于CAM激酶IV的底物并确定其各自 使用北部和免疫印迹和免疫细胞化学的本地化，以及 Camkkiks是否具有细胞类型或特定于组织的功能。