Molecular mechanism for the regulation of activation in calcium/calmodulin-dependent protein kinase II (CaMKII)

钙/钙调蛋白依赖性蛋白激酶 II (CaMKII) 激活调节的分子机制

• 批准号: 9431888
• 负责人: Moitrayee Bhattacharyya
• 金额: $ 9万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9431888
• 关键词: Address; Affect; Arrhythmia; Binding; Biochemical; Biological Assay; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcineurin; Calcium; Calcium/calmodulin-dependent protein kinase; Calmodulin; Cardiac; Cell physiology; Cells; Chordata; Complement; Complex; Computer Simulation; Defect; Development; Disease; Doctor of Philosophy; Down Syndrome; Drug Targeting; Electrophysiology (science); Enzymes; Exhibits; Frequencies; Glutamate Receptor; Goals; Heart failure; Holoenzymes; Homo; Human; Impairment; In Vitro; Kinetics; Label; Learning; Length; Lipid Bilayers; Long-Term Potentiation; Maintenance; Mammalian Cell; Mass Spectrum Analysis; Measurement; Measures; Membrane; Memory; Mental Health; Mentors; Methods; Microscopy; Modeling; Molecular; Molecular Conformation; Monitor; Mutation; N-Methyl-D-Aspartate Receptors; N-Methylaspartate; Neurons; Pathologic; Pathology; Phase; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiological; Play; Positioning Attribute; Preparation; Process; Property; Protein Isoforms; Protein-Serine-Threonine Kinases; Proteins; Protocols documentation; RNA Splicing; Reaction; Recruitment Activity; Regulation; Role; Signal Transduction; Signal Transduction Pathway; Site; Stimulus; Synapses; System; Techniques; Tissues; Training; Transducers; Variant; Withdrawal; Work; base; calmodulin-dependent protein kinase II; career development; cognitive development; insight; molecular dynamics; mutant; neuropathology; neurotransmission; overexpression; prevent; receptor; receptor binding; reconstitution; response; single molecule

PROJECT SUMMARY/ABSTRACT Calcium/calmodulin-dependent kinase II (CaMKII) is a Ser/Thr kinase, which plays a critical role in neuronal and cardiac signaling. Unregulated / perturbed activation of CaMKII leads to several pathological conditions, like severe impairment in learning and memory, cardiac arrhythmias and heart failure. Owing to the physiological importance of the activated state of CaMKII, it is obvious that the activation of CaMKII is tightly regulated in cells. My proposed project aims at understanding the molecular basis of this regulation from three perspectives: (1) It is interesting to note that the sequence of this Ca2+ responsive kinase is highly conserved in chordates, except an intrinsically disordered linker that connect the kinase domains to a central dodecameric/tetradecameric hub. Based on the length/composition of this linker, CaMKII is classified into four distinct isoforms and exhibits tissue-specific expression and specific subcellular localization. This intrinsically disordered linker is hypothesized to tune the ease of CaMKII activation in response to varying amplitude/frequency of Ca2+ signals. I will study the role of this intrinsically disordered linker in regulating the activation of CaMKII at a single molecule level. (2) We have recently discovered a unique phenomenon in CaMKII called subunit-exchange, whereby activated CaMKII subunits can exchange into unactivated CaMKII. This can be a mechanism to potentiate activation signal of CaMKII, long after the withdrawal of the initial Ca2+ stimulus. I will study the effect of blocking subunit exchange in cells and eventually I plan to establish a connection between spread of CaMKII activation signal and long-term potentiation/learning/memory. (3) Finally, I will reconstitute the regulatory circuit of CaMKII, including its membrane-associated interaction partners, such as the NMDA receptor, the dopaminergic D3 receptor and the phosphatases, on a supported membrane. Using, this simplistic model, I will study the effect of various signal inputs to this circuit to examine the regulatory constraints on CaMKII at the synapse. I need additional training in microscopy, mass spectrometry and electrophysiological measurements to address these questions. I have assembled a team of mentor and collaborators who will provide me with the technical and intellectual advice for my career development. My short-term (K99/early R00-phase) goal is to apply the newly learned techniques to answer pertinent questions related to the activation and regulation of CaMKII and its role in long-term potentiation, learning and memory. This will position me well to pursue my long- term goal to probe mental health development and maintenance from a molecular perspective. Specifically, I want to investigate the molecular mechanism underlying Down syndrome, focusing on the activation, regulation and downstream signaling effects of a Down syndrome critical region kinase DYRK1a, a promising drug target for Down syndrome neuropathologies.

项目总结/摘要 钙/钙调蛋白依赖性激酶II（CaMKII）是一种丝氨酸/苏氨酸激酶，在神经元和神经元细胞中起关键作用。 心脏信号CaMKII的不受调节/干扰的激活导致几种病理状况，如 严重损害学习和记忆、心律失常和心力衰竭。由于生理 由于CaMK II的活化状态的重要性，很明显，CaMK II的活化在细胞中受到严格调节。 我提议的项目旨在从三个角度了解这种调节的分子基础： (1)有趣的是，注意到这种Ca 2+响应激酶的序列在脊索动物中高度保守， 除了将激酶结构域连接到中心十二聚体/十四聚体的固有无序接头 枢纽基于该接头的长度/组成，CaMKII被分为四种不同的同种型，并表现出 组织特异性表达和特异性亚细胞定位。这种本质上无序的连接体被假设为 调节CaMKII激活的容易性以响应Ca 2+信号的变化幅度/频率。我会研究 这种内在无序的连接体在单分子水平上调节CaMKII活化的作用。(2)我们 我最近发现了CaMKII中一种独特的现象，称为亚基交换， 亚基可以交换成未活化的CaMKII。这可能是一种机制，以加强激活信号， CaMKII，在撤回初始Ca 2+刺激后很久。我将研究阻断亚基交换 最终，我计划建立CaMKII激活信号的传播与长期 增强/学习/记忆。(3)最后，我将重建CaMKII的调节回路，包括其 膜相关的相互作用伙伴，如NMDA受体，多巴胺能D3受体和 磷酸酶，在支持的膜上。利用这个简单的模型，我将研究各种信号的影响， 输入到这个电路，以检查突触处对CaMKII的调节约束。 我需要显微镜、质谱和电生理测量方面的额外培训， 回答这些问题。我已经组建了一个由导师和合作者组成的团队，他们将为我提供 为我的职业发展提供技术和知识方面的建议。我的短期（K99/R 00早期阶段）目标是 应用新学到的技术来回答有关激活和调节的相关问题， CaMKII及其在长时程增强、学习和记忆中的作用这将使我能够很好地追求我的长期- 长期目标是从分子角度探讨心理健康的发展和维持。我特别 我想研究唐氏综合征的分子机制，重点是激活，调节， 和唐氏综合征关键区激酶DYRK 1a的下游信号传导作用，一个有前途的药物靶点 唐氏综合症的神经病理学