Xu Ya-Wen, Lin Peng, Zheng Shu-Fa, Huang Wen, Lin Zhang-Ya, Shang-Guan Huang-Cheng, Lin Yuan-Xiang, Yao Pei-Sen, Kang De-Zhi
Department of Neurosurgery, Neurosurgical Research Institute, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
Department of Pain, The First Affiliated Hospital, Fujian Medical University, Fuzhou, China.
Front Neurol. 2021 Oct 1;12:713293. doi: 10.3389/fneur.2021.713293. eCollection 2021.
We test the hypothesis that lysine acetylation is involved in the metabolic process of glioma-associated seizures (GAS). We used label-free mass spectrometry-based quantitative proteomics to quantify dynamic changes of protein acetylation between gliomas with seizure (CA1 group) and gliomas without seizure (CA2 group). Furthermore, differences of acetyltransferase and deacetylase expression between CA1 and CA2 groups were performed by a quantitative proteomic study. We further classified acetylated proteins into groups according to cell component, molecular function, and biological process. In addition, metabolic pathways and protein interaction networks were analyzed. Regulated acetyltransferases and acetylated profiles were validated by PRM and Western blot. We detected 169 downregulated lysine acetylation sites of 134 proteins and 39 upregulated lysine acetylation sites of 35 proteins in glioma with seizures based on acetylome. We detected 407 regulated proteins by proteomics, from which ACAT2 and ACAA2 were the differentially regulated enzymes in the acetylation of GAS. According to the KEGG analysis, the upregulated acetylated proteins within the PPIs were mapped to pathways involved in the TCA cycle, oxidative phosphorylation, biosynthesis of amino acids, and carbon metabolism. The downregulated acetylated proteins within the PPIs were mapped to pathways involved in fatty acid metabolism, oxidative phosphorylation, TCA cycle, and necroptosis. Regulated ACAT2 expression and acetylated profiles were validated by PRM and Western blot. The data support the hypothesis that regulated protein acetylation is involved in the metabolic process of GAS, which may be induced by acetyl-CoA acetyltransferases.
我们检验了赖氨酸乙酰化参与胶质瘤相关性癫痫（GAS）代谢过程的假设。我们使用基于无标记质谱的定量蛋白质组学来量化有癫痫发作的胶质瘤（CA1组）和无癫痫发作的胶质瘤（CA2组）之间蛋白质乙酰化的动态变化。此外，通过定量蛋白质组学研究对CA1组和CA2组之间乙酰转移酶和脱乙酰酶的表达差异进行了分析。我们根据细胞成分、分子功能和生物学过程将乙酰化蛋白质进一步分类。此外，还分析了代谢途径和蛋白质相互作用网络。通过平行反应监测（PRM）和蛋白质免疫印迹法验证了调控的乙酰转移酶和乙酰化谱。基于乙酰化蛋白质组，我们在有癫痫发作的胶质瘤中检测到134种蛋白质的169个赖氨酸乙酰化位点下调以及35种蛋白质的39个赖氨酸乙酰化位点上调。通过蛋白质组学我们检测到407种受调控的蛋白质，其中乙酰辅酶A乙酰基转移酶2（ACAT2）和乙酰辅酶A酰基转移酶2（ACAA2）是GAS乙酰化过程中差异调控的酶。根据京都基因与基因组百科全书（KEGG）分析，蛋白质-蛋白质相互作用（PPI）中上调的乙酰化蛋白质被映射到参与三羧酸循环（TCA循环）、氧化磷酸化、氨基酸生物合成和碳代谢的途径。PPI中下调的乙酰化蛋白质被映射到参与脂肪酸代谢、氧化磷酸化、TCA循环和坏死性凋亡的途径。通过PRM和蛋白质免疫印迹法验证了调控的ACAT2表达和乙酰化谱。这些数据支持了蛋白质乙酰化调控参与GAS代谢过程这一假设，这可能是由乙酰辅酶A乙酰基转移酶诱导的。
