为探究大黄鱼在高温胁迫条件下的蛋白表达情况，实验应用串联质谱标签tandem mass tag (TMT)标记结合液相色谱-串联质谱(LC-MS/MS)技术对大黄鱼耐高温组和不耐高温组的肝脏组织进行蛋白组学分析。共鉴定到3 369个蛋白，其中有687个差异表达蛋白，包括281个上调蛋白和406个下调蛋白。用平行反应监测(parallel reaction monitoring, PRM)对随机挑选的13个蛋白进行验证，其结果与TMT标记的蛋白组学分析结果一致。随后，通过生物信息学分析发现，高温胁迫对大黄鱼的蛋白质折叠、能量代谢有显著影响，其中热休克蛋白70 (heat shock proteins 70, HSP70)、钙网蛋白(calreticulin, CRT)和葡萄糖调节蛋白(glucose-regulated protein, GRP)参与调节蛋白质的正确折叠，丙酮酸激酶(pyruvate kinase, PK)参与高温条件下的能量供给。由此推测HSP70、CRT、GRP和PK在大黄鱼应对高温胁迫时发挥着重要的作用。
In order to explore the changes in protein expression of Larimichthys crocea under high temperature stress, tandem mass tag (TMT) combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to perform proteomic analysis on the liver tissues between the high temperature tolerant group and the high temperature sensitive group of L. crocea. A total of 3 369 proteins were identified, of which 687 were differentially expressed proteins, including 281 up-regulated proteins and 406 down-regulated proteins. 13 randomly selected proteins were validated with parallel reaction monitoring (PRM) assays, which showed that they were consistent with the TMT-labeled proteomic analysis results. Subsequently, through bioinformatics analysis, it was found that high temperature stress had a significant impact on the protein folding and energy metabolism of L. crocea. Among them, heat shock protein 70(HSP70), calreticulin(CRT), and glucose-regulated protein(GRP) are involved in regulating the correct folding of proteins. Pyruvate kinase(PK) is involved in energy supply under high temperature conditions. It is speculated that HSP70, CRT, GRP and PK play an important role in the response of L. crocea to high temperature stress. This study is helpful to further analyze the high temperature adaptation mechanism of L. crocea, and provides reference for breeding high temperature resistant varieties of L. crocea.