Co-translational Regulation in the Vasculature of Organ Systems with Aging

项目总结/摘要 功能性蛋白质的生产是一个多步骤的过程， 多肽被酶促加工、折叠并组装成寡聚复合物。关键 成熟过程通过偶联mRNA翻译和新生蛋白协同发生， 在核糖体上成熟，由调节蛋白网络辅助。尽管经过数十年的 在基础研究中，我们对共翻译过程的理解主要由以下因素形成： 在原核和真核细胞模型中进行遗传操作，但在细胞中存在有限的数据 完整的哺乳动物生理系统。为了克服这一障碍，该项目旨在 全面了解新生蛋白质的共翻译调控，因为它们 在血管内皮细胞的血管生成信号通路内达到其功能状态， 心脏在老化。我们最近开发了一种方法，并测试了 同时分离存在于活跃翻译的多核糖体/mRNA内的蛋白质 心脏内皮内的复合物。最先进的多重标记和 这些蛋白质的定量已经允许鉴定一致的和不一致的 基于主动翻译mRNA和蛋白质谱的调节细胞特异性途径 与内皮应力有关。利用这一创新的进步，在“功能合作， 翻译组学”的方法，我们旨在研究内皮细胞内共翻译复合物的变化， 血管生成配体-受体和细胞-细胞相互作用的信号网络在老化的心脏。在 要求1 ，我们将定义一致调节的共翻译调节的变化， 新生儿和成年人的血管生成信号通路与年龄和性别。在 目的2 ，我们 将定义在不协调调节的血管生成中蛋白质的翻译后修饰 可能有助于心脏血管生成障碍的信号通路， 衰老最终，这些知识可以帮助开发更有效的手段， 诊断和治疗随着衰老发生的血管生成损伤， 心脏对心血管疾病和损伤的影响。

Project Summary/Abstract The production of functional proteins is a multistep process whereby newly synthesized polypeptides are enzymatically processed, folded, and assembled into oligomeric complexes. Key maturation processes occur co-translationally by coupling mRNA translation and nascent protein maturation on the ribosomes, assisted by a network of regulatory proteins. Despite decades of basic research, our understanding of co-translational processes is shaped predominantly by genetic manipulations in prokaryotic and eukaryotic cell models, but limited data exists in cells from intact mammalian physiological systems. To overcome this hurdle, this project aims to develop a comprehensive understanding of co-translational regulation of nascent proteins as they reach their functional state within the angiogenic signaling pathways of the endothelium in the heart during aging. We have recently developed an approach and tested the potential for simultaneous isolation of proteins present within the actively translating polyribosome/mRNA complexes within the endothelium of the heart. State-of-the-art multiplex labelling and quantification of these proteins has permitted the identification of concordant and discordant regulated cell-specific pathways based on actively translating mRNA and protein profiles associated with the endothelial stress. Leveraging this innovative advancement in “functional co- translatomics”, we aim to study the changes in co-translational complexes within endothelial cell angiogenic ligand-receptor and cell-cell interaction signaling networks within the aging heart. In Aim 1 , we will define changes in the co-translational regulation of concordantly regulated angiogenic signaling pathways during neonatal and adult life with aging and gender. In Aim 2 , we will define the post-translational modifications of proteins within discordantly regulated angiogenic signaling pathways which may contribute to the impairment angiogenesis in the heart during aging. Ultimately this knowledge could aid in the development of more effective means for diagnosing and treating the impairment of angiogenesis that occurs with aging and predisposes the heart to cardiovascular disease and injury.