广叶绣球菌子实体不同发育阶段的比较蛋白质组学分析

摘要

Protein expression at different developmental stages of Sparassis latifolia 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 2305 reliable proteins were identified using the Q-Exactive mass spectrometry and the ProteinPilot search engines,and 2219 of these proteins had a quantitative dimension.Of the quantitatively different proteins,104 were significantly up-regulated and 142 down-regulated at the early fruiting stage (80 days following inoculation),and 155 were significantly upregulated and 460 down-regulated at the fruiting stage (115 days following inoculation),compared with the primordium stage (60 days following inoculation),respectively.Data from gene ontology (GO) molecular functional analysis revealed that the differentially expressed proteins were mainly involved in catalytic activity,protein binding and hydrolase activity.Significantly enriched KEGG pathways included those related to carbon metabolism,biosynthesis of amino acids,ribosome pathways and glycolysis/gluconeogenesis.Furthermore,the numbers of differentially expressed proteins related to signal transmission and transcription factors were 27 and 7,respectively.iTRAQ-coupled 2D LC-MS/MS methodology was highly effective in isolating and identifying protein groups in S.latifolia,and could promote a better understanding of the developmental molecular mechanisms operative in this and other fungi.%利用同位素标记相对和绝对定量(isobaric tags for relative and absolute quantification,iTRAQ)标记结合二维液相色谱串联质谱(two-dimensional liquid chromatography tandem mass spectrometry,2D LC-MS/MS)技术,研究广叶绣球菌(Sparassis latifolia)原基期、幼菇期和成熟子实体阶段的差异表达蛋白质组.采用Q-Exactive质谱鉴定并经ProteinPilot软件搜库,对所获得的差异蛋白进行GO (gene ontology)、KEGG(kyoto encyclopedia of genes and genomes)和转录因子注释分析.结果共鉴定到可信蛋白2305个,其中2219个蛋白具有相对定量信息.与原基期相比,幼菇期显著上调蛋白104个,下调蛋白142个,子实体阶段显著上调蛋白155个,下调蛋白460个.GO分子功能提示这些差异性蛋白主要参与催化活性、蛋白结合和水解酶活性.KEGG代谢通路分析结果显示,差异蛋白主要涉及碳代谢、氨基酸合成、核糖体、糖酵解/糖衍生等代谢过程.差异蛋白中与信号传导和转录因子相关的蛋白数量分别为27个和7个.