Reversible protein acetylation and sirtuin function

描述（由申请人提供）：在过去几年中，越来越多的证据表明可逆乙酰化可能是控制蛋白质功能的主要调节机制。最近的蛋白质组学研究已经编目了数百种乙酰化蛋白的存在，暗示了几乎所有细胞生物化学方面的潜在调节作用。有趣的是，代谢酶构成乙酰化蛋白质的最大部分。蛋白质乙酰化的功能意义仅在少数例子中被揭示。因此，有巨大的需要，以确定蛋白质乙酰化的调节功能，无论是在生理结果的水平，并在分子机制的水平。该提议直接研究了可逆乙酰化是控制蛋白质功能的主要调节机制的中心假设。为了实现这些目标，我们将研究sirtuins的生化和生物学功能，NAD+依赖性蛋白脱乙酰酶，这涉及基因组的维持，代谢，细胞存活和寿命。具体的目标是集中在几个主要的代谢过程中，证明了sirtuins的直接调节作用，并阐明了位点特异性乙酰化如何影响酶的功能。采用机械酶学，定量质谱，新的高通量测定，代谢途径分析，结构生物学，酶调节，细胞培养模型和小鼠遗传学，这些研究将提供可逆的蛋白质乙酰化的功能意义的第一个全面的理解。这些结果有可能揭示一种以前未知的调节机制的重要性，并改变我们对代谢和年龄相关疾病的理解。 公共卫生相关性：新的证据表明，存在一种以前未知的细胞和代谢调节形式。该提案旨在研究这种调节机制的功能重要性以及一组涉及基因组维持，代谢，细胞存活和寿命的酶所发挥的作用。这些结果有可能改变我们对代谢和年龄相关疾病的理解，并产生新的治疗机会。

DESCRIPTION (provided by applicant): Over the last several years, accumulating evidence suggests that reversible acetylation may be a major regulatory mechanism for controlling protein function. Recent proteomic investigations have cataloged the existence of hundreds of acetylated proteins, implicating a potential regulatory role for nearly all facets of cellular biochemistry. Interestingly, metabolic enzymes constitute the largest portion of acetylated proteins. Only in a handful of examples has the functional significance of protein acetylation been revealed. Thus, there is tremendous need to determine the regulatory functions of protein acetylation, both at the level of physiological outcomes and at the level of molecular mechanism. This proposal directly investigates the central hypothesis that reversible acetylation is a major regulatory mechanism for controlling protein function. To accomplish these goals, we will investigate the biochemical and biological functions of sirtuins, NAD+dependent protein deacetylases, which are implicated in genome maintenance, metabolism, cell survival, and lifespan. The specific aims are focused on demonstrating a direct regulatory role for sirtuins in several major metabolic processes, and elucidating how site-specific acetylation affects enzyme function. Employing mechanistic enzymology, quantitative mass spectrometry, novel high-throughput assays, metabolic pathway analysis, structural biology, enzyme regulation, cell culture models, and mouse genetics, these studies will provide the first comprehensive understanding of the functional significance of reversible protein acetylation. The results have the potential to uncover the prominence of a previously-unknown regulatory mechanism and to transform how we understand metabolic and aged-related diseases. PUBLIC HEALTH RELEVANCE: New evidence suggests that a previously-unknown form of cellular and metabolic regulation exits. This proposal seeks to investigate the functional importance of this regulatory mechanism and the role played by a group of enzymes that are implicated in genome maintenance, metabolism, cell survival, and lifespan. The results have the potential to the transform how we understand metabolic and aged-related diseases, and to generate novel therapeutic opportunities.