用比较蛋白质组学研究烟曲霉的致病机制

发布时间：2018-05-26 17:24

  本文选题：烟曲霉 + 致病机制　； 参考：《吉林大学》2010年博士论文

【摘要】： 本研究以临床重要病原真菌烟曲霉(Aspergillus fumigatus)为材料,通过连续传代的方法,获得表型和致病力均发生明显变化的突变株。对野生株和突变株的胞内蛋白质、分泌蛋白质进行比较蛋白质组学研究,以及野生株的温度敏感性与致病性关系的研究,亦进行了不同菌株间胞内蛋白质的比较。胞内蛋白质图谱分析显示,有42个差异蛋白质点,通过质谱分析鉴定出9种。其中5种属于热休克蛋白家族(heat shock protein,Hsps),分别是Hsp90、Hsp88、Hsp70、HscA和Sti1。其他4种蛋白质分别为真核翻译起始因子EIF 4、核糖体60S亚基P0和2种肌动蛋白VIP 1和Act1。应用实时荧光定量分析技术对Hsp90、Hsp88、Sti1和EIF 4在基因水平的变化进行探讨,发现了3种热休克蛋白的mRNA水平和蛋白水平的表达一致。分泌蛋白质的研究中发现,野生株中有57种表达上调,突变株仅有11种上调。在不同温度(25℃和37℃)下,野生株胞内蛋白质有55个差异点,鉴定出6种:蛋白酶体调节亚单位Rpt3、热休克蛋白(Hsp82和Hsp BiP/Kar2)和3种“假想”蛋白质。上述实验发现热休克蛋白可能与致病性和温度变化有关。另外,还比较了来自临床和环境的6株烟曲霉的致病力差异,并对强毒株和弱毒株的胞内蛋白质进行了比较,选择其中49个差异点,鉴定出2种:菌丝体过氧化氢酶(CatB)和ATP合酶-β亚单位(ATP synthase F1,beta subunit)。并得到1种疑似Hsp70同系物。研究结果表明,热休克蛋白家族和过氧化氢酶、一些参与蛋白质合成、能量合成的蛋白质可能与烟曲霉的致病性相关。
[Abstract]:In this study, we used Aspergillus fumigatusas, an important pathogenic fungus, as the material, and obtained the mutants with obvious phenotype and pathogenicity by the method of continuous passage. The comparative proteomics of intracellular proteins and secretory proteins of wild and mutant strains and the relationship between temperature sensitivity and pathogenicity of wild strains were studied. The intracellular proteins of different strains were also compared. Analysis of intracellular protein map showed that there were 42 differential protein spots and 9 species were identified by mass spectrometry. Five of them belong to the heat shock protein family, Hsp90, Hsp88, Hsp70, HscA and Stil, respectively. The other four proteins were eukaryotic translation initiation factor EIF 4, ribosomal 60S subunit P0 and two actin VIP 1 and Act1, respectively. The changes of Hsp90 Hsp88-Sti1 and EIF _ 4 at gene level were studied by real-time fluorescence quantitative analysis. It was found that the mRNA level and protein level of three kinds of heat shock proteins were consistent. In the study of secreting proteins, 57 species of wild strains were up-regulated, while only 11 of mutant strains were up-regulated. At different temperatures (25 鈩，

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