应用iTRAQ结合2D LC-MS/MS技术分析草菇同核和异核菌丝蛋白质组

Analysis of the Proteomes in Volvariella volvacea Homokaryotic and Heterokaryotic Mycelia by iTRAQ-coupled 2-D LC-MS/MS

首次利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)技术结合二维液相色谱串联质谱对草菇(Volvariella volvacea)同核和异核菌丝蛋白质组进行研究。采用QExactive质谱鉴定并经MASCOT软件搜库,对所有鉴定蛋白进行主成分分析(principal component analysis,PCA)、层次聚类(hierarchical clustering)分析和GO(gene ontology)注释,并进行生物信息学分析。结果共获得2335个不同肽段,鉴定到1039个蛋白,其中1030个蛋白具有定量信息。在同核菌丝中显著上调蛋白83个,下调蛋白97个,表明iTRAQ标记技术结合二维液相色谱串联质谱可有效地分离和鉴定草菇菌丝蛋白质组。该研究结果为更深入全面地研究草菇菌丝蛋白质组学奠定基础。

Protein expression in homokaryotic and heterokaryotic mycelia of Volvariella volvacea has been investigated using isobaric tags for relative and absolute quantification（iTRAQ）-coupled 2D LC-MS/MS（two-dimensional liquid chromatography tandem mass spectrometry）.A total of 1039 proteins,including 2335 unique peptides,were identified using Q Exactive mass spectrometry and the MASCOT search engine,and 1030 of these had a quantitative dimension.Of the quantitatively different proteins,83 were significantly up-regulated and 97 down-regulated in homokaryotic,compared with heterokaryotic,mycelium.Data from principal component analysis,hierarchical clustering and gene ontology（GO） also revealed that iTRAQ-coupled 2D LC-MS/MS represented a highly effective method for isolating and identifying protein groups in V.volvacea mycelia,and would be useful in future analyses of the fungal proteome.