Kim Yonghyun, Nandakumar M P, Marten Mark R
Department of Chemical and Biochemical Engineering, University of Maryland Baltimore County (UMBC), 1000 Hilltop Circle, Baltimore, MD 21250, USA.
Fungal Genet Biol. 2007 Sep;44(9):886-95. doi: 10.1016/j.fgb.2006.12.001. Epub 2007 Jan 26.
The model filamentous fungus Aspergillus nidulans, when grown in a moderate level of osmolyte (+0.6M KCl), was previously found to have a significantly reduced cell wall elasticity (Biotech Prog, 21:292, 2005). In this study, comparative proteomic analysis via two-dimensional gel electrophoresis (2de) and matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) mass spectrometry was used to assess molecular level events associated with this phenomenon. Thirty of 90 differentially expressed proteins were identified. Sequence homology and conserved domains were used to assign probable function to twenty-one proteins currently annotated as "hypothetical." In osmoadapted cells, there was an increased expression of glyceraldehyde-3-phosphate dehydrogenase and aldehyde dehydrogenase, as well as a decreased expression of enolase, suggesting an increased glycerol biosynthesis and decreased use of the TCA cycle. There also was an increased expression of heat shock proteins and Shp1-like protein degradation protein, implicating increased protein turnover. Five novel osmoadaptation proteins of unknown functions were also identified.
模式丝状真菌构巢曲霉,当在中等水平的渗透剂(+0.6M KCl)中生长时,先前发现其细胞壁弹性显著降低(《生物技术进展》,21:292,2005)。在本研究中,通过二维凝胶电泳(2DE)和基质辅助激光解吸电离/飞行时间(MALDI-TOF)质谱进行比较蛋白质组学分析,以评估与该现象相关的分子水平事件。90个差异表达蛋白中有30个被鉴定出来。利用序列同源性和保守结构域为目前注释为“假定的”21种蛋白质赋予可能的功能。在渗透适应细胞中,3-磷酸甘油醛脱氢酶和醛脱氢酶的表达增加,而烯醇化酶的表达降低,这表明甘油生物合成增加,三羧酸循环的利用减少。热休克蛋白和Shp1样蛋白质降解蛋白的表达也增加,这意味着蛋白质周转增加。还鉴定出了5种功能未知的新型渗透适应蛋白。