Proteomics and Post-translational Modification

摘要 溶酶体是高度专门化和动态的细胞器，有助于维持蛋白质的动态平衡。 溶酶体也是细胞的信号中心。这些信号中的许多被传递到细胞核，并 最终调节基因转录和蛋白质产物的表达。这包括基因和 对溶酶体功能和生物发生至关重要的蛋白质。溶酶体基因的失调与 溶酶体系统功能障碍与衰老和年龄相关疾病有关，包括 阿尔茨海默氏症。蛋白质组学与定量翻译后修饰核心(蛋白质组学 核心)(核心B)将支持本PPG提案中的所有三个研究项目，以阐明 蛋白质组和蛋白质表达在衰老和阿尔茨海默病期间改变的机制。 我们将使用我们初步数据中描述的方法来测量(1)溶酶体的变化 蛋白质组，包括异构体和翻译后修饰，(2)翻译后修饰 转录上游的溶酶体相关信号事件，以及(3)组蛋白翻译后 溶酶体基因调控机制的修饰。蛋白质组学核心将利用我们的专业知识 和当前亚细胞细胞器蛋白质组学分析的能力，有针对性的蛋白质组特征 和蛋白质修饰状态(蛋白质形式)的量化，以及在信号方面的广泛专业知识 转导和组蛋白修饰。核心B的重点是提供可重现的量化和详细的 溶酶体和对溶酶体生物学调控至关重要的关键蛋白的特性 衰老和肌肉紧张症。这一水平的分子细节很重要，因为蛋白质属性，如异构体， 蛋白分解和翻译后修饰通常协同工作或在特定的层次中执行 它们的核心功能。总之，这些研究将提供无与伦比的详细描述，揭示 老年生物学和阿尔茨海默病中溶酶体功能、调节和生物发生的分子机制 疾病病理生理学。CORE B还将积极与其他核心和项目合作，创建第一个- 溶酶体蛋白质组和代谢组类衰老和疾病相关的溶酶体图谱(图谱) 小鼠的大脑皮质和海马体将与普通研究社区共享。

Abstract Lysosomes are highly specialized and dynamic cellular organelles that help maintain protein homeostasis. Lysosomes also act as a signaling center of the cell. Many of these signals are transduced to the nucleus and ultimately modulate transcription of genes and the expression of protein products. This includes genes and proteins that are essential to lysosome function and biogenesis. Dysregulation of lysosomal genes and the dysfunction of the lysosome system are associated with aging and age-associated diseases, including Alzheimer’s disease. The Proteomics and Quantitative Post-translational Modification core (proteomics core) (Core B) will support all three research projects in this PPG proposal to elucidate changes in the proteome and the mechanisms by which protein expression is altered during aging and Alzheimer’s disease. We will use methods as described in our preliminary data to measure changes in (1) the lysosomal proteome, including isoforms and post-translational modifications, (2) post translational modifications in lysosome-associated signaling events upstream of transcription, and (3) histone post-translational modifications in lysosomal gene regulatory mechanisms. The proteomics core will leverage our expertise and current capacity in the proteomic analysis of subcellular organelles, targeted proteomic characterization and quantitation of protein modification state (proteoforms), as well as extensive expertise in signal transduction and histone modifications. The focus of Core B is to provide reproducible quantitation and detailed characterization of the lysosome and crucial proteins that are essential to the regulation of lysosomal biology in aging and tauopathy. This level of molecular detail is important because protein attributes, such as isoforms, proteolysis, and post-translational modifications, often function in concert or in specific hierarchies to execute their core function. Together, these studies will provide unparalleled detailed characterization that reveal molecular mechanisms of lysosome function, regulation, and biogenesis in aging biology, and Alzheimer’s disease pathophysiology. Core B will also actively collaborate with other Cores and Projects to create a first-in- class Aging- and Tauopathy-associated Lysosomal atlas (ATLas) of the lysosomal proteome and metabolome for mouse cortex and hippocampus to be shared with the general research community.