Calcium/Calmodulin Activated Kinases in Smooth Muscle

平滑肌中钙/钙调蛋白激活的激酶

• 批准号: 6890456
• 负责人: HAROLD A SINGER
• 金额: $ 35.55万
• 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 2008-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6890456
• 关键词: RNA interference; calmodulin dependent protein kinase; carotid artery; cell migration; cell proliferation; enzyme activity; enzyme mechanism; enzyme structure; gene expression; immunocytochemistry; isozymes; laboratory rat; muscle cells; muscle contraction; phosphorylation; protein protein interaction; restenosis; tissue /cell culture; vascular smooth muscle

DESCRIPTION (provided by applicant): Ongoing work funded by this grant has focused on the role of Ca2+/calmodulin-dependent protein kinase II (CaMKII), a ubiquitous multifunctional serine/threonine kinase with unique structural and autoregulatory properties, in the control of vascular smooth muscle (VSM) contractile and growth responses. Our prior work in this area resulted in the biochemical and molecular characterization of CaMKII isozyme expression in VSM. Using this knowledge and experience, we have developed a number of specific immunochemical tools and molecular approaches that, for the first time, are capable of dissecting CaMKII activity and function in VSM. Recent studies have led to the general hypothesis that CaMKIIdelta2, the principal isoform expressed in cultured rat aortic VSM, has unique structural features that result in its coupling to nonreceptor tyrosine kinases and ERK1/2-dependent signaling pathways necessary for cell migration and growth. Biochemical and molecular approaches are proposed to: (Aim 1) test the requirement for and specificity of CaMKIIdelta2 isozymes in regulating Ca2+depedent activation of ERK1/2, migration, and proliferation in cultured VSM cells. (Aim 2) determine the functional significance of a novel CaMKIIdelta2/Fyn tyrosine kinase physical interaction with respect to CaMKll-dependent signaling and VSM cell migration and proliferation; (Aim 3) determine the effect of autophosphorylation in regulating CaMKlIdelta2 scaffolding functions and cellular function with respect to Ca2+dependent signaling, migration, and proliferation. Based on published analyses of CaMKII gene expression in VSM, we propose that a change in expression from CaMKII gamma-isozymes, which have been associated with control of VSM contraction, to the delta2-isozyme is an important component of the response to vascular injury and enables Ca2+-dependent control of VSM cell migration and growth. Aim 4 tests this hypothesis in the rat carotid artery injury and restenosis model. The proposed work includes the development of novel approaches and tools to address the function and regulation of specific CaMKII isozymes in arterial smooth muscle, focusing on cell migration and growth. Knowledge gained from these studies can potentially be applied to developing new therapeutic approaches for diseases that are known to involve vascular smooth muscle proliferation and migration such as atherosclerosis and restenosis.

描述（由申请人提供）：该基金资助的正在进行的工作集中在Ca 2 +/钙调素依赖性蛋白激酶II（CaMKII）的作用，CaMKII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，在控制血管平滑肌（VSM）收缩和生长反应中。我们以前在这方面的工作导致在VSM中的CaMKII同工酶表达的生化和分子表征。利用这些知识和经验，我们已经开发了一些特定的免疫化学工具和分子方法，第一次，能够解剖在VSM中的CaMKII活性和功能。最近的研究已经导致了一般的假设，即CaMKII δ 2，在培养的大鼠主动脉VSM中表达的主要亚型，具有独特的结构特征，导致其偶联到非受体酪氨酸激酶和ERK 1/2依赖的信号转导通路细胞迁移和生长所必需的。本研究拟从生物化学和分子生物学的角度探讨：（1）在培养的VSM细胞中，CaMK Ⅱ δ 2同工酶在调节依赖Ca ~（2+）的ERK 1/2激活、迁移和增殖中的需要和特异性。(Aim 2）确定新的CaMKII δ 2/Fyn酪氨酸激酶物理相互作用在CaMKII依赖性信号传导和VSM细胞迁移和增殖方面的功能意义;（目的3）确定自磷酸化在调节CaMKII δ 2支架功能和细胞功能中的作用，所述细胞功能与Ca 2+依赖性信号传导、迁移和增殖有关。基于已发表的分析，在VSM中的CaMKII基因表达，我们提出，从CaMKII γ-同工酶，这已与VSM收缩控制，δ 2-同工酶的表达变化是一个重要的组成部分，血管损伤的反应，使钙依赖性的VSM细胞迁移和生长的控制。目的4在大鼠颈动脉损伤再狭窄模型中验证这一假说。拟议的工作包括开发新的方法和工具，以解决动脉平滑肌中特定CaMKII同工酶的功能和调节，重点是细胞迁移和生长。从这些研究中获得的知识可以潜在地应用于开发新的治疗方法，用于已知涉及血管平滑肌增殖和迁移的疾病，如动脉粥样硬化和再狭窄。