Molecular Regulation of CaM Kinase II Activity

CaM 激酶 II 活性的分子调控

• 批准号: 6513664
• 负责人: SONNY Thomas ABRAHAM
• 金额: $ 15.63万
• 依托单位: CAMPBELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6513664
• 关键词: biological signal transduction; calcium ion; calmodulin dependent protein kinase; cell cycle; enzyme activity; enzyme induction /repression; fluorescence microscopy; immunocytochemistry; immunoprecipitation; ion transport; phosphomonoesterases; phosphorylation; protein structure function; vascular smooth muscle

DESCRIPTION (provided by applicant): Changes in intracellular calcium ion concentrations ([Ca2+li) has been linked to the progression of the cell division cycle of mammalian cells. The mechanism by which this universal second messenger regulates such a complex process is unclear; however, many of its actions are most likely mediated by calcium-sensitive protein kinases and phosphatases. Recent work on the identification and characterization of calcium/calmodulin-dependent protein kinase 11 (CaM kinase 11) in rat aortic vascular smooth muscle (VSM) cells indicates that this enzyme is responsive to changes in [Ca2+]i and so, may play a fundamental role in processes such as cell division. It is well accepted that cellular signaling events are regulated by phosphorylation and dephosphorylation of critical cellular proteins. There is also compelling evidence that kinases and phosphatases are themselves targets of such phosphorylation/dephosphorylation events. Further, the complex activation/inactivation steps of CaM kinase 11 would uniquely lend itself to regulation by specific cellular phosphatases. An emerging paradigm of signal transduction networks is the sequestration of kinase/phosphatase complexes with committed downstream cellular function. This proposal seeks to identify and characterize phosphatases that regulate CaM kinase 11 activity in cultured VSM cells. This will be achieved by determining how CaM kinase 11 activity changes when endogenous phosphatases are inhibited through pharmacological and physiological manipulations. These phosphatases will be characterized by immunological and activity assays and, evidence for kinase/phosphatase signalling complexes will be sought through colocalization and cell fractionation studies. The applicant's preliminary work suggests that CaM kinase 11 plays a prominent role in the cell division process of VSM cells. Such a role takes on added significance when one considers that unregulated proliferation of VSM cells is a fundamental process in the development of atherosclerotic lesions and subsequent heart disease. The characterization of novel cellular processes would hasten the development of more selective pharmacological and genetic tools that may be used to better manage or reverse the disease process.

描述（由申请人提供）：细胞内钙离子浓度的变化（[CA2+LI）与哺乳动物细胞的细胞分裂周期的进展有关。这个通用的第二信使调节这种复杂过程的机制尚不清楚。但是，其许多作用很可能是由钙敏感蛋白激酶和磷酸酶介导的。大鼠主动脉血管平滑肌（VSM）细胞中钙/钙调蛋白依赖性蛋白激酶11（CAM激酶11）的鉴定和表征的最新工作表明，该酶对[Ca2+] i的变化有反应[Ca2+] i，因此可能在细胞分裂等过程中起着基本作用。众所周知，细胞信号传导事件受到关键细胞蛋白的磷酸化和去磷酸化的调节。还有令人信服的证据表明，激酶和磷酸酶本身是这种磷酸化/去磷酸化事件的靶标。此外，CAM激酶11的复合激活/灭活步骤将独特地通过 特定的细胞磷酸酶。信号转导网络的新兴范式是 具有下游细胞功能的激酶/磷酸酶复合物的固相。 该建议旨在识别和表征调节培养的VSM细胞中CAM激酶11活性的磷酸酶。这将通过确定CAM激酶11活性在通过药理和生理操纵抑制内源性磷酸酶时如何变化来实现。这些磷酸酶将以免疫学和活性测定的特征，以及通过共定位和细胞分馏研究寻求激酶/磷酸酶信号传导复合物的证据。申请人的初步工作表明，CAM激酶11在VSM细胞的细胞分裂过程中起着重要作用。当人们认为VSM细胞不管制的增殖是动脉粥样硬化病变和随后的心脏病的发展中，这种作用具有额外的意义。新颖的细胞过程的表征将加快可以用来改善的更有选择性的药理学和遗传工具的发展 管理或扭转疾病过程。