In-depth analysis of the Sirtuin 5-regulated mouse brain malonylome and succinylome using library-free data-independent acquisitions.
In-depth analysis of the Sirtuin 5-regulated mouse brain malonylome and succinylome using library-free data-independent acquisitions.
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翻译后修饰（PTM）动态调节蛋白质和生物学途径，通常通过多种PTM的组合作用。赖氨酸残基靶向各种PTM，包括丙二酰化和琥珀酰化。然而，PTM在数据采集和处理过程中为基于质谱的蛋白质组学提供了特定的挑战。因此，使用数据独立采集（DIA）的新颖和创新的工作流程可确保可靠的PTM鉴定，精确的位点定位以及准确和稳健的无标记定量。在这项研究中，我们提出了一个强大的方法，结合抗体为基础的富集与全面的DIA收购和光谱库免费数据处理使用directDIA（Spectronaut）。相同的DIA数据可用于生成光谱库，全面识别和量化PTM，减少富集样品量和所需的采集时间，同时提供全自动化的工作流程。我们分析了野生型和Sirtuin 5（SIRT5）基因敲除小鼠的大脑，发现并定量了466个丙二酰化和2211个琥珀酰化肽。SIRT5调控通过靶向164个丙二酰化位点和578个琥珀酰化位点重塑了酰基组。受影响的途径包括碳水化合物和脂质代谢，突触囊泡周期和神经退行性疾病。我们发现了48个常见的SIRT5调节的丙二酰化和琥珀酰化位点，表明潜在的PTM串扰。这种创新和高效的工作流程提供了对小鼠大脑赖氨酸丙二酰组和琥珀酰组的更深入的了解。
Post-translational modifications (PTMs) dynamically regulate proteins and biological pathways, typically through the combined effects of multiple PTMs. Lysine residues are targeted for various PTMs, including malonylation and succinylation. However, PTMs offer specific challenges to mass spectrometry-based proteomics during data acquisition and processing. Thus, novel and innovative workflows using data-independent acquisition (DIA) ensure confident PTM identification, precise site localization, and accurate and robust label-free quantification. In this study, we present a powerful approach that combines antibody-based enrichment with comprehensive DIA acquisitions and spectral library-free data processing using directDIA (Spectronaut). Identical DIA data can be used to generate spectral libraries and comprehensively identify and quantify PTMs, reducing the amount of enriched sample and acquisition time needed, while offering a fully automated workflow. We analyzed brains from wild-type and Sirtuin 5 (SIRT5)-knock-out mice, and discovered and quantified 466 malonylated and 2211 succinylated peptides. SIRT5 regulation remodeled the acylomes by targeting 164 malonylated and 578 succinylated sites. Affected pathways included carbohydrate and lipid metabolisms, synaptic vesicle cycle, and neurodegenerative diseases. We found 48 common SIRT5-regulated malonylation and succinylation sites, suggesting potential PTM crosstalk. This innovative and efficient workflow offers deeper insights into the mouse brain lysine malonylome and succinylome.
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SIRT5 使代谢相关蛋白脱酰并减轻 ob/ob 小鼠的肝脂肪变性
影响因子：
11.1
作者：
Du Y;Hu H;Qu S;Wang J;Hua C;Zhang J;Wei P;He X;Hao J;Liu P;Yang F;Li T;Wei T
通讯作者：
Wei T
影响因子：
4
作者：
Doll, Sophia;Burlingame, Alma L.
通讯作者：
Burlingame, Alma L.
影响因子：
4.4
作者：
Burger, Thomas
通讯作者：
Burger, Thomas
DOI：
10.3390/antiox6020025
发表时间：
2017-04-05
期刊：
Antioxidants (Basel, Switzerland)
影响因子：
--
作者：
Gao J;Wang L;Liu J;Xie F;Su B;Wang X
通讯作者：
Wang X
赖氨酸丙二酰化的蛋白质组学和生化研究表明其在线粒体功能和脂肪酸氧化中与丙二酸尿症相关的调节作用。
影响因子：
7
作者：
Colak, Gozde;Pougovkina, Olga;Zhao, Yingming
通讯作者：
Zhao, Yingming