Quantification of Mitochondrial Acetylation Dynamics Highlights Prominent Sites of Metabolic Regulation
Quantification of Mitochondrial Acetylation Dynamics Highlights Prominent Sites of Metabolic Regulation
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DOI：
10.1074/jbc.m113.483396
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发表时间：
2013-09-06
影响因子：
4.8
通讯作者：
Pagliarini, David J.
中科院分区：
文献类型：
--
作者：
Still, Amelia J.;Floyd, Brendan J.;Pagliarini, David J.
赖氨酸乙酰化正在迅速成为调节线粒体新陈代谢的关键翻译后修饰。然而，从数千个通过质谱学鉴定的调控位点中区分出来，并阐明这些修饰如何改变酶的功能仍然是主要的挑战。在这里，我们进行了多路定量质谱分析，以测量小鼠肝脏线粒体乙酰化蛋白质组在营养状态急性和慢性变化时的变化，并将这些数据集与我们的预测SIRT3靶点纲要整合在一起。这些分析突出了具有动态乙酰化位点的线粒体蛋白质的子集，包括乙酰辅酶A乙酰基转移酶1(Acat1)，这是一种对多种代谢途径至关重要的酶。我们进行了体外生物化学和分子模拟，证明了Acat1的乙酰化通过破坏辅酶A的结合而降低其活性。总之，我们的数据揭示了一个重要的调控乙酰化的新靶点，并为研究选择线粒体蛋白乙酰化位点在介导急性和慢性代谢转变中的作用提供了基础。
Lysine acetylation is rapidly becoming established as a key post-translational modification for regulating mitochondrial metabolism. Nonetheless, distinguishing regulatory sites from among the thousands identified by mass spectrometry and elucidating how these modifications alter enzyme function remain primary challenges. Here, we performed multiplexed quantitative mass spectrometry to measure changes in the mouse liver mitochondrial acetylproteome in response to acute and chronic alterations in nutritional status, and integrated these data sets with our compendium of predicted Sirt3 targets. These analyses highlight a subset of mitochondrial proteins with dynamic acetylation sites, including acetyl-CoA acetyltransferase 1 (Acat1), an enzyme central to multiple metabolic pathways. We performed in vitro biochemistry and molecular modeling to demonstrate that acetylation of Acat1 decreases its activity by disrupting the binding of coenzyme A. Collectively, our data reveal an important new target of regulatory acetylation and provide a foundation for investigating the role of select mitochondrial protein acetylation sites in mediating acute and chronic metabolic transitions.