CALCIUM/CALMODULIN ACTIVATED KINIASES IN SMOOTH MUSCLE

平滑肌中钙/钙调蛋白激活的激酶

• 批准号: 6638340
• 负责人: HAROLD A SINGER
• 金额: $ 31万
• 依托单位: ALBANY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-07-01 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6638340
• 关键词: calcium flux; calmodulin dependent protein kinase; enzyme activity; enzyme mechanism; enzyme structure; enzyme substrate; isozymes; laboratory rabbit; laboratory rat; protein purification; protein sequence; transfection; vascular smooth muscle

DESCRIPTION (Verbatim from applicant's application): A long-term goal of this laboratory has been to elucidate the function of specific protein kinases in the control of vascular smooth muscle (VSM) contractile and growth responses. Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) is a ubiquitous multifunctional serine/threonine kinase with unique structural and autoregulatory properties. The general goal of this proposal is to understand to what extent CaM Kinase II medicates the diverse consequences of intracellular Ca2+ signals in VASM. Combined pharmacological and molecular approach employing recombinant replication defective adenoviruses to infect cultured VSM cells with specific active CaM kinase II subunits of inactive mutants will be used to the test the following hypothesis: (AIM 1) CaM kinase II is an intermediate in the Ca2+-dependent activation of non-receptor tyrosine kinases and the MAP kinase signaling pathway in VSM cells. (AIM 2) CaM kinase II isozymes are involved in the Ca2+-dependent regulation of VSM migration and integrin-dependent signaling; (AIM 3) CaM Kinase II is involved in the regulation of tonic contractile function and protein tyrosine phosphorylation responses in intact arterial smooth muscle. Detailed structural information pertaining to CaM kinase II subunit and holoenzyme has also raised interesting questions regarding to the relationships between CaM kinase II isozyme structure and its unique autoregulatory properties or subcellular localization. AIM 4 proposes experiments to define specific amino acid residues that are (auto) phosphorylated within CaM kinase II following activation of VSM cells by physiological stimuli, and the relationships between phosphorylation of these sites and modulation of CaM kinase II activity. Finally, in AIM 5, we propose to determine the structural basis for specific subcellular targeting of CaM kinase II isozymes in VSM and to identify binding or targeting proteins involved in modulating CaM kinase II localization or activity. The proposed research addresses questions relating to one mechanism (activation of CaM kinase II) through which intracellular Ca2+ signals may regulate fundamental cellular processes such as gene transcription, cell motility, and proliferation. Specifically, the research will lead to a more complete understanding of the mechanisms for controlling VSM cell migration and contraction, processes that are involved in pervasive vascular diseases including atherosclerosis, restenosis, hypertension and vasospasm.

描述（来自申请人的申请）：这是一个长期目标 实验室已经阐明了特定蛋白激酶在 控制血管平滑肌（VSM）的收缩和生长反应。 Ca 2 +/钙调蛋白依赖性蛋白激酶II（CaM激酶II）是一种普遍存在的 多功能丝氨酸/苏氨酸激酶具有独特的结构和 自动调节特性本提案的总体目标是了解 CaM激酶II在多大程度上治疗了 VASM细胞内Ca ~（2+）信号。结合药理学和分子生物学 使用重组复制缺陷型腺病毒感染 培养的VSM细胞具有特异性活性CaM激酶II亚基， 突变体将用于测试以下假设：（AIM 1）CaM激酶 II是非受体酪氨酸的Ca 2+依赖性活化的中间体 激酶和MAP激酶信号通路。(AIM 2）CaM激酶 II同工酶参与了VSM迁移的Ca ~（2+）依赖性调节， 整合素依赖性信号传导;（AIM 3）CaM激酶II参与整合素依赖性信号传导。 紧张性收缩功能和蛋白酪氨酸磷酸化的调节 完整动脉平滑肌的反应。详细结构信息 关于CaM激酶II亚基和全酶的研究也引起了人们的兴趣， 关于CaM激酶II同工酶之间关系的问题 结构及其独特的自动调节特性或亚细胞定位。 AIM 4提出了定义特定氨基酸残基的实验， 在VSM细胞活化后，CaM激酶II内的（自动）磷酸化 生理刺激，以及这些磷酸化之间的关系， 位点和调节CaM激酶II活性。最后，在AIM 5中，我们建议 确定CaM特异性亚细胞靶向的结构基础 VSM中的激酶II同工酶并鉴定结合或靶向蛋白 参与调节CaM激酶II定位或活性。拟议 研究解决了与一种机制有关的问题（钙调素的激活 激酶II），通过细胞内Ca 2+信号可以调节基本的 细胞过程，如基因转录、细胞运动和 增殖具体来说，这项研究将导致一个更完整的 了解控制VSM细胞迁移的机制， 收缩，参与广泛性血管疾病的过程 包括动脉粥样硬化、再狭窄、高血压和血管痉挛。