Journal of Proteomics ( IF 2.8 ) Pub Date : 2020-09-03 , DOI: 10.1016/j.jprot.2020.103967 Yan Zhao 1 , Gang Yang 1 , Ningyuan Wu 2 , Xiaojuan Cao 1 , Jian Gao 1
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Fatty acid desaturate 2 (Fads2) is associated with many chronic diseases. Nevertheless, comprehensive researches on its role have not been performed. We here conducted an integrated analysis of long-chain polyunsaturated fatty acid (LC-PUFA) metabolism of fads2-deletion zebrafish (fads2−/−) by transcriptomics, proteomics and phosphoproteomics. Compared with wild type zebrafish (WT), fads2−/− showed significantly higher contents of hepatic linoleic acid (all-cis-9,12-C18:2), α-linolenic acid (all-cis-9,12,15-C18:3) and docosapetaenoic acid (all-cis-7,10,13,16,19-C22:5), and lower contents of γ-linolenic acid (all-cis-6,9,12-C18:3), stearidonic acid (all-cis-6,9,12,15-C18:4) and docosahexaenoic acid (all-cis-4,7,10,13,16,19-C22:6), accompanied by an increased n-6/n-3 PUFA level. In total, we identified 1608 differentially expressed genes (DEGs), 209 differentially expressed proteins (DEPs) and 153 differentially expressed phosphorylated proteins (DEPPs) with 190 sites between fads2−/− and WT. Transcriptome and proteome analysis simultaneously aggregated these DEGs and DEPs into LC-PUFA synthesis and PPAR signaling pathways. Further interaction network analysis of the DEPPs showed that spliceosome and protein processing in endoplasmic reticulum pathway were critical groups. Additionally, we determined seven highly phosphorylated kinases and a highly expressed phosphatase in fads2−/− zebrafish. These results give insights into the mechanism by which fads2 affects metabolic disease occurrence, and provide datasets for target selections for human disease treatment.
Significance
Balanced LC-PUFA composition was deeply associated with body health, while changes of LC-PUFAs usually induced serious diseases such as cardiovascular disease, type 2 diabetes and inflammatory disease. Fatty acid desaturase 2 (Fads2), subordinating to the fatty acid desaturase protein family, catalyzes the first desaturation reaction in LC-PUFA synthesis. Although Fads2 is associated with many chronic diseases including metabolic abnormalities, type 2 diabetes and obesity, comprehensive researches on its role have not been performed.
On the basis of the integrated transcriptome, proteome and phosphoproteome analysis, we identified that fads2 was critical for maintaining body LC-PUFA homeostasis. Moreover, the crucial pathways including PPAR signaling pathway, spliceosome and protein processing in endoplasmic reticulum pathway, and candidate kinase targets associated with LC-PUFA metabolism were determined. These findings will contribute to the revealing of the mechanism and supply possible datasets for target selection for human disease treatment.
中文翻译：
综合转录组和磷酸化蛋白质组分析表明，fads2 对于维持身体 LC-PUFA 稳态至关重要。
去饱和脂肪酸 2 (Fads2) 与许多慢性疾病有关。然而，尚未对其作用进行全面的研究。我们在这里通过转录组学、蛋白质组学和磷酸蛋白质组学对fads2缺失斑马鱼 ( fads2 -/- )的长链多不饱和脂肪酸 (LC-PUFA) 代谢进行了综合分析。与野生型斑马鱼 (WT) 相比，fads2 -/-显示肝脏亚油酸 (all-cis-9,12-C18:2)、α-亚麻酸 (all-cis-9,12,15-C18:3) 和二十二碳五烯酸 (all-cis-7 ,10,13,16,19-C22:5), γ-亚麻酸 (all-cis-6,9,12-C18:3), 硬脂酸 (all-cis-6,9,12 ,15-C18:4) 和二十二碳六烯酸 (all-cis-4,7,10,13,16,19-C22:6)，伴随着增加的 n-6/n-3 PUFA 水平。我们总共鉴定了 1608 个差异表达基因 (DEG)、209 个差异表达蛋白 (DEP) 和 153 个差异表达磷酸化蛋白 (DEPP)，其中 190 个位点位于fads2 -/-和WT。转录组和蛋白质组分析同时将这些 DEG 和 DEP 聚合到 LC-PUFA 合成和 PPAR 信号通路中。DEPPs的进一步相互作用网络分析表明，内质网途径中的剪接体和蛋白质加工是关键组。此外，我们在fads2 -/-斑马鱼中确定了七种高度磷酸化的激酶和一种高度表达的磷酸酶。这些结果深入了解了fads2影响代谢疾病发生的机制，并为人类疾病治疗的目标选择提供了数据集。
意义
平衡的 LC-PUFA 组成与身体健康密切相关，而 LC-PUFA 的变化通常会诱发心血管疾病、2 型糖尿病和炎症性疾病等严重疾病。脂肪酸去饱和酶 2 (Fads2) 属于脂肪酸去饱和酶蛋白家族，催化 LC-PUFA 合成中的第一个去饱和反应。尽管 Fads2 与代谢异常、2 型糖尿病和肥胖等多种慢性疾病有关，但尚未对其作用进行全面研究。
在综合转录组、蛋白质组和磷酸化蛋白质组分析的基础上，我们发现fads2对维持身体 LC-PUFA 稳态至关重要。此外，还确定了包括 PPAR 信号通路、内质网通路中的剪接体和蛋白质加工在内的关键通路，以及与 LC-PUFA 代谢相关的候选激酶靶点。这些发现将有助于揭示机制并为人类疾病治疗的目标选择提供可能的数据集。
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