Reversible protein acetylation and sirtuin function

描述（由申请人提供）：在过去的几年中，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，暗示了细胞生物化学几乎所有方面的潜在调节作用。有趣的是，代谢酶构成了乙酰化蛋白的最大部分。只有在少数例子中，蛋白质乙酰化的功能意义才被揭示出来。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。该建议直接调查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 （NAD+依赖性蛋白去乙酰化酶）的生化和生物学功能，这些功能与基因组维持、代谢、细胞存活和寿命有关。具体目标集中在证明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的功能。利用机制酶学、定量质谱法、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，这些研究将首次全面了解可逆蛋白乙酰化的功能意义。这些结果有可能揭示一个以前未知的调节机制的重要性，并改变我们对代谢和衰老相关疾病的理解。

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.