Molecular Mechanisms of Protein Arginylation

蛋白质精氨酸化的分子机制

• 批准号: 9068168
• 负责人: Anna S Kashina
• 金额: $ 31.3万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9068168
• 关键词: Acidic Amino Acids; Address; Affect; Affinity; Aging; Biological; Catalysis; Cause of Death; Cell Extracts; Cellular Metabolic Process; Chemistry; Complex; Data; Development; Disease; Enzymes; Event; Goals; Health; Heart Diseases; Human; Kinetics; Lead; Malignant Neoplasms; Mediating; Molecular; Morphogenesis; N-terminal; Nerve Degeneration; Peptides; Perception; Physiological; Physiological Processes; Physiology; Play; Post-Translational Protein Processing; Process; Property; Protein Isoforms; Proteins; Proteolysis; Proteome; Reaction; Regulation; Series; Side; Signal Pathway; Site; Specificity; Stimulus; Substrate Specificity; Testing; Therapeutic; United States; base; cell motility; cofactor; design; human disease; in vitro activity; in vivo; insight; novel; novel therapeutics; protein function; research study; response; synthetic peptide

DESCRIPTION (provided by applicant): Posttranslational modification of proteins in signal pathways is one of the most pivotal control points in vivo for the activation, inactivation, and degradation of factors governing nearly all physiological processes. The goal of this proposal is to define the molecular mechanism and in vivo functions of protein arginylation, a very poorly understood post-translational modification that affects an estimated 25% of the cellular proteome. Arginylation plays a key role in cell migration and developmental morphogenesis, and is implicated in major events of cell metabolism, physiology, and human disease. Our preliminary data strongly suggest that arginylation regulates its protein targets through rapid and reversible modulation of their activity and protein interactions and constitutes a truly essential and dynamic biological regulator, however virtually nothing is known about the substrate site specificity and the molecular mechanisms of this reaction. This proposal will address the mechanisms of the arginyl transfer enzyme (ATE1) and the regulation of its substrate specificity through an integrated approach that will provide the first mechanistic insights into this enigmatic posttranslational modification. We propose the following specific aims: (1) To determine site specificity and properties of N- terminal arginylation; (2) To test the mechanisms of arginylation at internal protein sites in intact proteins; and (3) To test the effect of cellular components on he rate and site specificity of arginylation by different ATE1 isoforms and modulate its activity in vivo. Together, the proposed studies will elucidate the molecular mechanism of a novel posttranslational modification with major biological significance. These studies will ultimately enable differential modulation of ATE1 activity and biological targeting, essential for exploring is therapeutic potential in critical arginylation-dependent human conditions, including heart disease, cancer, neurodegeneration, and aging. The results of our studies will open new possibilities of functional arginylation analysis and targeted manipulation of arginylation of key proteins during essential processes in normal physiology and disease.

描述（由申请人提供）：信号通路中蛋白质的翻译后修饰是体内控制几乎所有生理过程的因子的激活、失活和降解的最关键控制点之一。该提案的目标是定义蛋白质精氨酸化的分子机制和体内功能，这是一种人们知之甚少的翻译后修饰，影响了估计 25% 的细胞蛋白质组。精氨酰化在细胞迁移和发育形态发生中起着关键作用，并与细胞代谢、生理学和人类疾病的重大事件有关。我们的初步数据强烈表明，精氨酸化通过快速和稳定的方式调节其蛋白质靶标。 它们的活性和蛋白质相互作用的可逆调节，并构成真正重要的动态生物调节剂，但实际上对底物位点特异性和该反应的分子机制一无所知。该提案将通过综合方法解决精氨酰转移酶 (ATE1) 的机制及其底物特异性的调节，该方法将为这一神秘的机制提供第一个机制见解。 翻译后修饰。我们提出以下具体目标：（1）确定N-末端精氨酰化的位点特异性和性质； (2) 测试完整蛋白质内部蛋白质位点精氨酸化的机制； (3)测试细胞成分对不同ATE1异构体精氨酰化的速率和位点特异性的影响并调节其体内活性。总之，拟议的研究将阐明具有重大生物学意义的新型翻译后修饰的分子机制。这些研究最终将实现 ATE1 活性和生物靶向的差异调节，对于探索关键的精氨酰化依赖性人类疾病（包括心脏病、癌症、神经退行性疾病和衰老）的治疗潜力至关重要。我们的研究结果将为功能性精氨酸化分析和正常生理和疾病的重要过程中关键蛋白质精氨酸化的靶向操作开辟新的可能性。