Defining the landscape and mechanisms of protein redox regulation during aging

项目摘要：哺乳动物组织从事的特殊生理活动是通过 蛋白质半胱氨酸残基的ROS可逆性修饰。ROS管理着无数的 生物学过程，而ROS和氧化还原信号的失调是最长的假设基础之一 随着年龄增长，组织生理性衰退的原因。尽管氧化还原监管的普遍重要性 组织特定的生理学和哺乳动物的衰老，一直缺乏关于特定的信息 解释体内这些过程的分子基础的蛋白质修饰。 我们最近开发了一种质谱学(MS)技术，用于第一次全面和定量 小鼠半胱氨酸氧化还原蛋白质组的体内定位。我们目前的目标是定义景观和 半胱氨酸氧化网络的机制，是年龄相关组织病理和寿命的基础。我们的 初步数据显示，所有老年人都发生了半胱氨酸氧化网络的根本性重构 组织，其中许多网络映射到已建立的与疾病相关的蛋白质家族。我们将测试 协调氧化还原调节支配蛋白质的空间组织、组装和功能的假说 已知与年龄相关疾病相关的网络。根据更多的初步数据，我们将 检验这样一种假设，即有力地延长寿命和健康的干预措施会导致系统重塑 蛋白质半胱氨酸氧化网络。 为了检验这些假设，我们将追求以下具体目标。在目标1中，我们将系统地确定 半胱氨酸氧化在协调这些蛋白质复杂网络组装中所起的作用。此外，对于 已经与衰老疾病相关的两个优先蛋白质网络，我们将定义半胱氨酸的作用 氧化对相应的生物功能和寿命的影响。在目标2中，我们将氧化还原受监管的网络 饮食限制对延年益寿的调控干预将决定那些延年益寿的半胱氨酸 通过应用平行氧化还原，在整个进化过程中，跨性别和保守的氧化网络 线虫衰老模型和Dr.的蛋白质组学分析我们将进一步确定其机制和 新发现的α-酮戊二酸代谢的氧化还原调节靶点的氧化还原调节的代谢后果， 我们推测，它在DR介导的代谢调节中起着核心作用。总而言之，我们建议 首次定义了定义组织老化和DR的蛋白质组范围的氧化还原调控图景。 依赖于寿命的提升。这些目标的成功实现将确定一种模式的机制 ROS的生物调节长期以来一直与衰老有关，但几十年来一直难以捉摸。

PROJECT SUMMARY: Mammalian tissues engage in specialized physiology that is regulated through reversible modification of protein cysteine residues by reactive oxygen species (ROS). ROS regulate a myriad of biological processes, and dysregulation of ROS and redox signaling is one of the longest postulated underlying causes of physiological decline of tissues with age. Despite the widespread importance of redox regulation of tissue-specific physiology and mammalian aging, there is a persistent lack of information regarding the specific protein modifications that explain the molecular basis for these processes in vivo. We recently developed a mass spectrometric (MS) technology for the first comprehensive and quantitative mapping of the mouse cysteine redox proteome in vivo. Our current objective is to define the landscape and mechanisms of cysteine oxidation networks that underlie age-dependent tissue pathology and lifespan. Our preliminary data demonstrate a fundamental remodeling of cysteine oxidation networks occurs in all aged tissues, and many of these networks map to established disease-relevant protein families. We will test the hypothesis that coordinated redox regulation governs spatial organization, assembly, and function of protein networks already known to be relevant to age-related disease. Building on additional preliminary data, we will test the hypothesis that interventions that robustly extend lifespan and healthspan exert systematic remodeling of protein cysteine oxidation networks. To test these hypotheses, we will pursue the following specific aims. In Aim 1 we will systematically determine the role that cysteine oxidation plays in coordinating these protein complex network assemblies. Moreover, for two priority protein networks already associated with diseases of aging, we will define the role of cysteine oxidation on corresponding biological function and lifespan. In Aim 2 we will redox regulated networks that are regulated by the longevity promoting intervention of dietary restriction will determine those pro-longevity cysteine oxidation networks that are conserved across sexes and throughout evolution by applying parallel redox proteomics analyses in models of C. elegans aging and DR. We will additionally determine the mechanisms and metabolic consequences of redox regulation of newfound redox regulated targets of α-ketoglutarate metabolism, which we hypothesize play a central role in DR-mediated metabolic regulation. Taken together, we propose to define, for the first time, the proteome-wide redox regulatory landscape that defines tissue aging and DR- dependent promotion of lifespan. Successful completion of these Aims will define mechanisms for a mode of biological regulation by ROS that has long been associated with aging, but for decades has remained elusive.