Structural and functional studies of protein kinase C regulation

蛋白激酶C调节的结构和功能研究

• 批准号: 10598531
• 负责人: Tatyana I. Igumenova
• 金额: $ 30.58万
• 依托单位: TEXAS A&M AGRILIFE RESEARCH
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598531
• 关键词: Address; Affinity; Alzheimer' s Disease; Basic Science; Binding; Binding Sites; Biochemical; Biophysics; Cells; Chemicals; Complex; Coupled; Data; Detection; Development; Diabetes Mellitus; Diglycerides; Ensure; Enzymes; Eukaryotic Cell; Event; Exhibits; Family; Funding; HIV/AIDS; Health; Heart Diseases; Human; Human Pathology; Hydrophobicity; In Vitro; Isoenzymes; Knowledge; Ligands; Lipids; Malignant Neoplasms; Membrane; Membrane Lipids; Membrane Proteins; Molecular; Molecular Conformation; Mutation; Outcome; Pharmacologic Substance; Phenotype; Phosphatidylinositols; Phospholipids; Phosphotransferases; Play; Process; Protein Kinase C; Protein Kinase C Inhibitor; Proteins; Regulation; Research Proposals; Resolution; Role; Signal Pathway; Signal Transduction; Signaling Protein; Structural Models; Structure; System; Tertiary Protein Structure; Testing; Therapeutic Research; Tumor Promoters; Tumor Suppressor Proteins; biophysical techniques; design; experimental study; falls; insight; molecular dynamics; novel; pharmacologic; phosphatidylinositol phosphate, PtdIns(4,5)P2; prevent; protein activation; protein kinase modulator; recruit; response; simulation; tumor progression; two-dimensional

Protein Kinases C (PKC) define a family of lipid-activated kinases that are key effectors of phosphoinositide signaling – a major intracellular signaling pathway of eukaryotic cells. Consistent with this central activity, dysregulation of PKC signaling is implicated in cancer progression, cardiac disease, diabetes, and Alzheimer's disease. Because of the fundamental role of PKC in signal transduction, the development of modulators of PKC activity – both for therapeutic and basic research purposes – is widely recognized as one of the major challenges in the field. Addressing these challenges requires an atomic-level understanding of PKC control – in particular, how lipids regulate PKC activity. This subject defines a major gap in understanding of PKC regulatory mechanisms. The central objective of this research proposal is to decipher the mechanism of the key event that triggers activation of PKC – its interaction with diacylglycerol (DAG). It is currently unknown how DAG is recognized and captured in membranes by the conserved homology 1 (C1) domains of PKC. To address this critical gap in knowledge, a synergistic combination of advanced solution NMR approaches, atomistic molecular dynamics simulations, and biochemical strategies will be applied to address the following Specific Aims: (1) to understand the molecular basis of diacylglycerol recognition by C1 domains, (2) to decipher precisely how C1 domains are initially recruited to membranes, and (3) to determine how tandem C1 domains execute a coincidence detection of DAG and anionic phospholipids. Insights into the DAG-sensing mechanism obtained from the proposed studies will have impact that extends beyond the already large problem of PKC regulation. The atomic-level information regarding C1-DAG interactions will be directly applicable to five other large families of signaling proteins that rely on C1 domains for regulation of their DAG- dependent activities. This information will also facilitate the design of pharmacological agents for rational modulation of PKC function by direct targeting of its C1 domains.

蛋白激酶 C (PKC) 定义了脂质激活激酶家族，它们是磷酸肌醇的关键效应子 信号传导——真核细胞的主要细胞内信号传导途径。与这一中心活动相一致， PKC 信号传导失调与癌症进展、心脏病、糖尿病和阿尔茨海默病有关 疾病。由于 PKC 在信号转导中的基本作用，调节剂的开发 PKC 活性——无论是用于治疗目的还是基础研究目的——被广泛认为是主要的活性之一。 该领域的挑战。应对这些挑战需要对 PKC 控制有原子级的理解 – 特别是脂质如何调节 PKC 活性。该主题定义了对 PKC 理解的主要差距 监管机制。本研究计划的中心目标是破译该机制 触发 PKC 激活的关键事件——它与二酰基甘油 (DAG) 的相互作用。目前尚不清楚 DAG 如何被 PKC 的保守同源 1 (C1) 结构域识别并捕获在膜中。到 解决这一知识上的关键差距，先进的解决方案核磁共振方法的协同组合， 原子分子动力学模拟和生化策略将应用于解决以下问题 具体目标：(1) 了解 C1 结构域识别二酰基甘油的分子基础，(2) 准确破译 C1 结构域最初如何招募到膜上，以及 (3) 确定串联 C1 如何 结构域执行 DAG 和阴离子磷脂的符合检测。深入了解 DAG 传感 从拟议的研究中获得的机制将产生超出已经很大的影响 PKC调节问题。有关 C1-DAG 相互作用的原子级信息将直接 适用于依赖 C1 结构域调节其 DAG 的其他五个信号蛋白大家族 依赖活动。这些信息也将有助于药物制剂的合理设计 通过直接靶向其 C1 结构域来调节 PKC 功能。