Comprehensive analysis of posttranslational protein modifications in aging of subcellular compartments
Comprehensive analysis of posttranslational protein modifications in aging of subcellular compartments
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DOI：
10.1038/s41598-020-64265-0
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发表时间：
2020-05-05
影响因子：
4.6
通讯作者：
Glomb, Marcus A.
中科院分区：
文献类型：
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作者：
Baldensperger, Tim;Eggen, Michael;Glomb, Marcus A.
通过氧化、糖化和酰化进行的酶促和非酶促翻译后蛋白质修饰是炎症、表观遗传学改变和蛋白质稳态下降等衰老标志的关键调节机制。在这项研究中，使用小鼠队列来监测衰老过程中翻译后修饰的变化。开发并验证了从小鼠肝脏中提取组蛋白、胞浆蛋白和线粒体蛋白的方案。总共，使用 HPLC-MS/MS 定量了亚细胞区室蛋白质中的 6 种赖氨酸酰化结构、7 种高级糖基化终产物、6 种氧化应激标记物和瓜氨酸化。蛋氨酸亚砜、乙酰化、甲酰化和瓜氨酸化是最丰富的修饰。组蛋白是在衰老过程中在所有亚细胞区室中积累的非凡的高修饰和非酶修饰。组蛋白乙酰化在年轻和年老动物中产生 350 至 305 μmol/mol 亮氨酸当量，与此相比，酰化、糖化和瓜氨酸化等修饰分别提高了乙酰化的 43%、20% 和 18%。另一方面，选定的氧化应激标记物与年龄相关的增加高达 150%。本研究中测量的数据和模式对于进一步的研究是强制性的，并将极大地促进对衰老分子机制的理解。
Enzymatic and non-enzymatic posttranslational protein modifications by oxidation, glycation and acylation are key regulatory mechanisms in hallmarks of aging like inflammation, altered epigenetics and decline in proteostasis. In this study a mouse cohort was used to monitor changes of posttranslational modifications in the aging process. A protocol for the extraction of histones, cytosolic and mitochondrial proteins from mouse liver was developed and validated. In total, 6 lysine acylation structures, 7 advanced glycation endproducts, 6 oxidative stress markers, and citrullination were quantitated in proteins of subcellular compartments using HPLC-MS/MS. Methionine sulfoxide, acetylation, formylation, and citrullination were the most abundant modifications. Histone proteins were extraordinary high modified and non-enzymatic modifications accumulated in all subcellular compartments during the aging process. Compared to acetylation of histone proteins which gave between 350 and 305 mu mol/mol leucine equivalents in young and old animals, modifications like acylation, glycation, and citrullination raised to 43%, 20%, and 18% of acetylation, respectively. On the other hand there was an age related increase of selected oxidative stress markers by up to 150%. The data and patterns measured in this study are mandatory for further studies and will strongly facilitate understanding of the molecular mechanisms in aging.