高糖微环境下小鼠晶状体上皮细胞中赖氨酸乙酰化的蛋白质组学分析

Proteomic analysis of lysine acetylation in mouse lens epithelial cells under high-glucose microenvironment

目的探讨高糖微环境下小鼠晶状体上皮细胞中赖氨酸乙酰化生物学过程和功能。方法采用小鼠晶状体上皮细胞系α-TN4建立正常处理组(葡萄糖5.5 mmol/L)和高糖处理组(葡萄糖45 mmol/L)。利用稳定同位素标记氨基酸标记2组细胞,使用泛乙酰化抗体富集,高分辨率液相色谱串联质谱(LC-MS/MS)检测和生物信息学数据挖掘。基因本体(GO)、亚细胞定位、真核直系同源组(KOG)、GO富集、京都基因与基因组百科全书(KEGG)富集、结构域富集和复合物富集分析小鼠晶状体上皮细胞中的乙酰化生物学过程和功能。结果在高糖微环境下,晶状体上皮细胞中乙酰化分别在分子功能、细胞成分、生物学过程、KOG和亚细胞定位中存在上调或下调变化;功能富集显示乙酰化主要参与和影响物质和能量代谢以及线粒体生物学过程。结论高糖诱导的赖氨酸乙酰化参与和改变多种生物功能、通路、结构域和蛋白质复合物,进而改变众多蛋白的表达及其功能。

Objective To explore the role of acetylation in mouse lens epithelial cells under high-glucose microenvironment. Methods A mouse lens epithelial cell line α-TN4 was treated with a normal level of glucose(5.5 mmol/L) and a high level of glucose(45 mmol/L). Both groups were labelled with stable isotope labeling by amino acids(SILAC) and analyzed using pan-acetylated antibody enrichment,high-resolution liquid chromatography-mass spectrometry detection,and bioinformatics data mining. Gene ontology(GO),subcellular localization,clusters of eukaryotic orthologous groups(KOG),GO enrichment,Kyoto encyclopeida of genes and genomes(KEGG) enrichment,domain enrichment,and complex enrichment were used to analyze acetylome in mouse lens epithelial cells. Results Under high-glucose microenvironment,there were up-and down-regulated changes in molecular functions,cellular components,biological processes,KOG and subcellular localization in lens epithelial cells. Functional enrichment showed that acetylation participated and affected the metabolism of substances and energy,as well as biological processes in the mitochondria. Conclusion High glucoseinduced lysine acetylation modifies the expressions and functions of many proteins by participating in and changing various biological functions,pathways,domains,and protein complexes.