Reversible protein acetylation and sirtuin function

描述(申请人提供)：在过去的几年里，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控机制。最近的蛋白质组学研究已经编目了数百种乙酰化蛋白质的存在，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，代谢酶构成了乙酰化蛋白质的最大部分。只有在少数几个例子中，蛋白质乙酰化的功能意义才被揭示。因此，无论是在生理结果水平上还是在分子机制水平上，都迫切需要确定蛋白质乙酰化的调节功能。这一建议直接调查了可逆乙酰化是控制蛋白质功能的主要调控机制这一中心假设。为了实现这些目标，我们将研究sirtuins的生化和生物学功能，sirtuins是NAD+依赖的蛋白脱乙酰酶，它与基因组维持、新陈代谢、细胞存活和寿命有关。具体目的是展示sirtuins在几个主要代谢过程中的直接调节作用，并阐明位点特异性乙酰化如何影响酶功能。这些研究将利用机械酶学、定量质谱学、新型高通量分析、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，首次全面了解可逆蛋白质乙酰化的功能意义。这一结果有可能揭示一种以前不为人知的调节机制，并改变我们对代谢和衰老相关疾病的理解。 与公共健康相关：新的证据表明，存在一种以前未知的细胞和代谢调节形式。这项建议旨在调查这种调节机制的功能重要性，以及一组与基因组维持、新陈代谢、细胞存活和寿命有关的酶所起的作用。这些结果有可能改变我们对新陈代谢和与衰老相关的疾病的理解，并创造新的治疗机会。

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.