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

项目摘要/摘要 功能蛋白质的生产是一个多步骤的过程，新合成的蛋白质 多肽经过酶处理、折叠和组装成低聚复合体。钥匙 成熟过程通过将信使核糖核酸翻译和新生蛋白偶联来共同翻译。 在核糖体上的成熟，在调控蛋白网络的帮助下。尽管几十年来 基础研究，我们对协译过程的理解主要是由 原核和真核细胞模型中的遗传操作，但细胞中存在的数据有限 来自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节，因为它们 在血管内皮细胞的血管生成信号通路中达到其功能状态 心脏在老化过程中。我们最近开发了一种方法，并测试了 同时分离主动翻译的多聚核糖体/mRNA中存在的蛋白质 心脏内皮细胞内的复合体。最先进的多用途标签和 对这些蛋白质的定量可以识别出一致性和不一致性 基于主动翻译信使核糖核酸和蛋白质谱的细胞特异性调控通路 与血管内皮细胞应激有关。利用这一创新的进步，在“功能联合 我们的目标是研究内皮细胞内共翻译复合体的变化。 老化心脏内的血管生成配体-受体和细胞-细胞相互作用信号网络。在……里面 目标1 ，我们将定义协调监管的协译监管的变化 新生儿和成人生命中血管生成信号通路随年龄和性别的变化。在……里面 目标2 ，我们 将定义不协调调节的血管生成中蛋白质的翻译后修饰 心脏血管生成障碍的信号转导途径 衰老。最终，这些知识可以帮助开发更有效的方法 增龄性和易感性血管生成障碍的诊断和治疗 心脏到心血管疾病和损伤。

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.