Protein kinase C-delta actions in cardiomyocytes

蛋白激酶 C-delta 在心肌细胞中的作用

基本信息

批准号：
8462652
负责人：
Susan F Steinberg
金额：
$ 37.39万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-08-15 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8462652
关键词：
Agonist Apoptosis Biochemical CREB1 gene Cardiac Cardiac Myocytes Cardiomyopathies Cell Fraction Cells Cytosol Development Docking Enzymatic Biochemistry Enzymes Etiology Event Evolution Funding Future GTP-Binding Proteins Generations Genetically Engineered Mouse Goals Growth Factor Growth Factor Receptors HDAC5 gene Heart failure Hypertrophy In Vitro Ischemia Lead Link Lipids Membrane Membrane Lipids Methods Modeling Molecular Molecular Conformation Mutagenesis Oxidative Stress Pathogenesis Peptide Library Peptides Pharmacologic Substance Phosphorylation Phosphotransferases Post-Translational Protein Processing Process Property Protein Kinase Protein-Serine-Threonine Kinases Proteins RNA Interference Reaction Reactive Oxygen Species Receptor Activation Recruitment Activity Reperfusion Injury Reperfusion Therapy Signal Transduction Site Specificity Stimulus Substrate Specificity Testing Treatment Efficacy Troponin I Tyrosine Tyrosine Phosphorylation Tyrosine Phosphorylation Site adapter protein adenoviral-mediated base cofactor delta protein design improved in vivo inhibitor/antagonist novel overexpression p66(ShcA) protein prevent protein kinase C-delta protein kinase D protein kinase modulator public health relevance receptor response small molecule therapeutic target

项目摘要

DESCRIPTION (provided by applicant): Protein kinase C4 (PKC4) is a serine/threonine kinase implicated in the pathogenesis of cardiac remodeling and reperfusion injury. PKC4 activation is generally attributed to receptor-driven, lipid cofactor-dependent mechanisms that anchor PKC4 in its active conformation to membranes. This allosteric model assumes that PKC4 activity is an inherent/immutable property of the enzyme that is not altered by the activation process. However, studies in HL77680 implicate PKC4 autophosphorylation and PKC4 tyrosine phosphorylation by Src as post-translational modifications that lead to functionally important changes in PKC4 phosphorylation of physiologically relevant target substrates. We also show that a tyrosine phosphorylated form of PKC4 accumulates in the cytosolic fraction of cardiomyocytes subjected to oxidative stress as a lipid-independent enzyme that is poised to phosphorylate substrates throughout the cell, not just on lipid membranes. Other studies show that PKC4 activation leads to the phosphorylation of a range of effector proteins with varied (and potentially opposing) effects on cellular remodeling. Studies in this renewal will focus on the PKC4 autophosphorylation and tyrosine phosphorylation mechanisms that dictate PKC4's cellular actions and can be targeted therapeutically to prevent adverse cardiac remodeling. Specific Aim I will use biochemical methods to identify the sites and consequences of PKC4 autophosphorylation and tyrosine phosphorylation by Src. The goal is to identify post-translational modifications that regulate PKC4 catalytic activity, and particularly its substrate specificity. Specific aim II will use RNAi silencing and adenoviral-mediated overexpression strategies to determine whether distinct molecular forms of PKC4, that accumulate in a stimulus-specific manner, regulate different effector responses. Studies in Specific Aim III will focus on the mechanism underlying the cardioprotective actions of peptide modulators of PKC4 translocation. The focus is on whether these compounds exert 'off-target' actions to prevent docking interactions required for regulatory phosphorylations on the enzyme. Specific aim IV will examine the molecular requirements for PKC4- dependent cardiac remodeling in vivo in genetically-engineered mice. The long-term goals are [1] to distinguish the cardiac actions of the allosterically-activated form of PKC4 that accumulates in response to growth factor receptor activation versus the tyrosine phosphorylated form of PKC4 that accumulates during oxidative stress and [2] to test the hypothesis that only certain molecular forms of PKC4 recruit effectors that promote adverse cardiac remodeling and ischemia/reperfusion injury. This information would provide the basis for the development of novel pharmaceuticals designed to prevent adverse cardiac remodeling by selectively inhibiting the cellular actions of specific molecular forms of PKC4 (or post-translational modifications on the enzyme).

描述（由申请人提供）：蛋白激酶C4（PKC4）是一种丝氨酸/苏氨酸激酶，与心脏重塑和再灌注损伤的发病机制有关。 PKC4 激活通常归因于受体驱动的脂质辅因子依赖性机制，该机制将 PKC4 以其活性构象锚定在膜上。该变构模型假设 PKC4 活性是酶的固有/不变特性，不会因激活过程而改变。然而，HL77680 的研究表明 PKC4 自磷酸化和 Src 引起的 PKC4 酪氨酸磷酸化是翻译后修饰，导致生理相关靶底物的 PKC4 磷酸化发生功能上重要的变化。我们还表明，酪氨酸磷酸化形式的 PKC4 在遭受氧化应激的心肌细胞的胞质部分中积累，作为一种脂质独立酶，它能够磷酸化整个细胞内的底物，而不仅仅是脂质膜上的底物。其他研究表明 PKC4 激活会导致一系列效应蛋白磷酸化，对细胞重塑产生不同（且可能相反）的影响。此次更新的研究将重点关注 PKC4 自身磷酸化和酪氨酸磷酸化机制，这些机制决定 PKC4 的细胞作用，并且可以作为治疗目标来预防不良心脏重塑。具体目标我将使用生化方法来鉴定 Src 引起的 PKC4 自磷酸化和酪氨酸磷酸化的位点和后果。目标是确定调节 PKC4 催化活性的翻译后修饰，特别是其底物特异性。具体目标 II 将使用 RNAi 沉默和腺病毒介导的过表达策略来确定以刺激特异性方式积累的不同分子形式的 PKC4 是否调节不同的效应器反应。 Specific Aim III 的研究将重点关注 PKC4 易位肽调节剂的心脏保护作用的机制。重点是这些化合物是否发挥“脱靶”作用，以防止酶上调节磷酸化所需的对接相互作用。具体目标 IV 将检查基因工程小鼠体内 PKC4 依赖性心脏重塑的分子需求。长期目标是 [1] 区分响应生长因子受体激活而积累的 PKC4 变构激活形式与在氧化应激过程中积累的 PKC4 酪氨酸磷酸化形式的心脏作用，并 [2] 测试假设只有某些分子形式的 PKC4 会募集促进不良心脏重塑和缺血/再灌注损伤的效应子。该信息将为开发新型药物提供基础，这些药物旨在通过选择性抑制 PKC4 特定分子形式（或酶的翻译后修饰）的细胞作用来预防不良心脏重塑。

项目成果

期刊论文数量（16）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Protein kinase D1 autophosphorylation via distinct mechanisms at Ser744/Ser748 and Ser916.

蛋白激酶 D1 通过 Ser744/Ser748 和 Ser916 的不同机制进行自磷酸化。

DOI：
发表时间：
2009-01-23
期刊：
影响因子：
0
作者：
Rybin, Vitalyi O;Guo, Jianfen;Steinberg, Susan F
通讯作者：
Steinberg, Susan F

The C2 Domain and Altered ATP-Binding Loop Phosphorylation at Ser³⁵⁹ Mediate the Redox-Dependent Increase in Protein Kinase C-δ Activity.

C2 结构域和 Ser 的 ATP 结合环磷酸化改变介导蛋白激酶 C-δ 活性的氧化还原依赖性增加。

DOI：
发表时间：
2015-05
期刊：
影响因子：
5.3
作者：
Gong, Jianli;Yao, Yongneng;Zhang, Pingbo;Udayasuryan, Barath;Komissarova, Elena V;Chen, Ju;Sivaramakrishnan, Sivaraj;Van Eyk, Jennifer E;Steinberg, Susan F
通讯作者：
Steinberg, Susan F

Protein kinase C-{delta} regulates the subcellular localization of Shc in H2O2-treated cardiomyocytes.

蛋白激酶 C-{delta} 调节 H2O2 处理的心肌细胞中 Shc 的亚细胞定位。