Defining the landscape and mechanisms of protein redox regulation during aging

项目概述：哺乳动物组织参与通过以下方式调节的专门生理学 通过活性氧（ROS）可逆修饰蛋白质半胱氨酸残基。ROS调节着无数 ROS和氧化还原信号的失调是生物过程中最长的假设之一， 随着年龄的增长，组织的生理衰退的原因。尽管氧化还原调节的广泛重要性， 组织特异性生理学和哺乳动物衰老，一直缺乏关于特定的 蛋白质修饰解释了这些过程在体内的分子基础。 我们最近开发了一种质谱（MS）技术， 小鼠体内半胱氨酸氧化还原蛋白质组图谱。我们目前的目标是定义景观， 半胱氨酸氧化网络是年龄依赖性组织病理学和寿命的基础。我们 初步数据表明，半胱氨酸氧化网络的基本重塑发生在所有老年人中， 这些网络中的许多映射到已建立的疾病相关蛋白质家族。我们将测试 协同氧化还原调节控制蛋白质空间组织、组装和功能的假说 已知与年龄相关疾病有关的网络。根据初步数据，我们将 测试这一假设，即干预措施，有力地延长寿命和健康寿命发挥系统性重塑 蛋白质半胱氨酸氧化网络。 为了验证这些假设，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合物网络组装中的作用。而且对于 两个优先蛋白质网络已经与衰老疾病，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，我们将氧化还原调节网络， 受饮食限制的长寿促进干预的调节将决定那些促长寿的半胱氨酸 氧化网络是保守的跨性别和整个进化，通过应用平行的氧化还原 蛋白质组学分析在C. elegans老化和DR。我们还将确定机制， 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， 我们假设其在DR介导的代谢调节中起核心作用。综上所述，我们建议 首次定义了定义组织衰老和DR的蛋白质组范围内的氧化还原调节景观， 寿命的依赖性增长。成功地完成这些目标将确定一种机制， 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.