Institute of Chemistry, Food Chemistry, Martin-Luther-University Halle-Wittenberg, Kurt-Mothes-Str. 2, 06120, Halle/Saale, Germany.
Clinic for Heart Surgery, Martin-Luther-University Halle-Wittenberg, Ernst-Grube Str. 40, 06120, Halle/Saale, Germany.
Sci Rep. 2020 May 5;10(1):7596. doi: 10.1038/s41598-020-64265-0.
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 µmol/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.
氧化、糖基化和酰化等酶促和非酶促的蛋白质翻译后修饰是衰老标志（如炎症、表观遗传学改变和蛋白质稳态下降）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、细胞质和线粒体蛋白的方案。使用 HPLC-MS/MS 共定量了亚细胞区室中蛋白质的 6 种赖氨酸酰化结构、7 种晚期糖基化终产物、6 种氧化应激标志物和瓜氨酸化。甲硫氨酸亚砜、乙酰化、甲酰化和瓜氨酸化是最丰富的修饰。组蛋白蛋白修饰异常丰富，非酶促修饰在衰老过程中在所有亚细胞区室中积累。与年轻和老年动物之间组蛋白蛋白的乙酰化产生 350 到 305 µmol/mol 亮氨酸等价物相比，酰化、糖基化和瓜氨酸化等修饰分别增加到 43%、20%和 18%。另一方面，一些选定的氧化应激标志物的含量随年龄的增长增加了 150%。本研究中测量的数据和模式是进一步研究的必要条件，将极大地促进对衰老分子机制的理解。
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