An acetylome peptide microarray reveals specificities and deacetylation substrates for all human sirtuin isoforms
An acetylome peptide microarray reveals specificities and deacetylation substrates for all human sirtuin isoforms
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DOI：
10.1038/ncomms3327
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发表时间：
2013-09-01
影响因子：
16.6
通讯作者：
Steegborn, Clemens
中科院分区：
文献类型：
--
作者：
Rauh, David;Fischer, Frank;Steegborn, Clemens
Sirtuin 酶通过目标蛋白中乙酰赖氨酸的脱乙酰作用来调节新陈代谢和衰老过程。已知超过 6,800 个哺乳动物乙酰化位点，但大多数 Sirtuin 异构体很少有靶标，这阻碍了我们对 Sirtuin 功能的理解。在这里，我们描述了一个显示 6,802 个人类乙酰化位点的肽微阵列系统，用于对其脱乙酰酶修饰的平行表征。通过该系统获得的所有七种人类沉默调节蛋白的脱乙酰化数据揭示了所有沉默调节蛋白同种型（包括Sirt4）的同种型特异性底物偏好和脱乙酰化底物候选。我们确认苹果酸脱氢酶蛋白是 Sirt3 底物，并表明过氧化还原蛋白 1 和高迁移率 B1 族蛋白分别在确定的位点被 Sirt5 和 Sirt1 脱乙酰化，使它们可能成为新的体内底物。我们的微阵列平台能够对生理乙酰化位点进行并行研究，所提供的脱乙酰化数据为鉴定所有人类去乙酰化酶的新型底物提供了令人兴奋的资源。
Sirtuin enzymes regulate metabolism and aging processes through deacetylation of acetyl-lysines in target proteins. More than 6,800 mammalian acetylation sites are known, but few targets have been assigned to most sirtuin isoforms, hampering our understanding of sirtuin function. Here we describe a peptide microarray system displaying 6,802 human acetylation sites for the parallel characterisation of their modification by deacetylases. Deacetylation data for all seven human sirtuins obtained with this system reveal isoform-specific substrate preferences and deacetylation substrate candidates for all sirtuin isoforms, including Sirt4. We confirm malate dehydrogenase protein as a Sirt3 substrate and show that peroxiredoxin 1 and high-mobility group B1 protein are deacetylated by Sirt5 and Sirt1, respectively, at the identified sites, rendering them likely new in vivo substrates. Our microarray platform enables parallel studies on physiological acetylation sites and the deacetylation data presented provide an exciting resource for the identification of novel substrates for all human sirtuins.