Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Science Park 904, 1098XH Amsterdam, The Netherlands.
Microbiology (Reading). 2011 Aug;157(Pt 8):2297-2307. doi: 10.1099/mic.0.049395-0. Epub 2011 May 20.
The ability of Candida albicans to switch from yeast to hyphal growth is essential for its virulence. The walls and especially the covalently attached wall proteins are involved in the primary host-pathogen interactions. Three hyphal induction methods were compared, based on fetal calf serum, the amino sugar N-acetylglucosamine (GlcNAc) and the mammalian cell culture medium Iscove's modified Dulbecco's medium (IMDM). GlcNAc and IMDM were preferred, allowing stable hyphal growth over a prolonged period without significant reversion to yeast growth and with high biomass yields. We employed Fourier transform-MS combined with a (15)N-metabolically labelled reference culture as internal standard for relative quantification of changes in the wall proteome upon hyphal induction. A total of 21 wall proteins were quantified. Our induction methods triggered a similar response characterized by (i) a category of wall proteins showing strongly increased incorporation levels (Als3, Hwp2, Hyr1, Plb5 and Sod5), (ii) another category with strongly decreased levels (Rhd3, Sod4 and Ywp1) and (iii) a third one enriched for carbohydrate-active enzymes (including Cht2, Crh11, Mp65, Pga4, Phr1, Phr2 and Utr2) and showing only a limited response. This is, to our knowledge, the first systematic, quantitative analysis of the changes in the wall proteome of C. albicans upon hyphal induction. Finally, we propose new wall-protein-derived candidates for vaccine development.
白色念珠菌从酵母形态到菌丝形态转换的能力对于其毒力至关重要。细胞壁，特别是共价连接的细胞壁蛋白，参与了主要的宿主-病原体相互作用。我们比较了三种菌丝诱导方法，分别基于胎牛血清、氨基糖 N-乙酰氨基葡萄糖（GlcNAc）和哺乳动物细胞培养基 Iscove 的改良 Dulbecco 培养基（IMDM）。GlcNAc 和 IMDM 是首选方法，可在较长时间内稳定诱导菌丝生长，而不会显著逆转回酵母生长状态，并且具有较高的生物量产量。我们采用傅里叶变换-MS 结合（15）N 代谢标记的参考培养物作为内部标准，用于相对定量分析菌丝诱导后细胞壁蛋白质组的变化。共定量了 21 种细胞壁蛋白。我们的诱导方法引发了相似的反应，其特征是：（i）一类细胞壁蛋白显示出强烈增加的掺入水平（Als3、Hwp2、Hyr1、Plb5 和 Sod5）；（ii）另一类蛋白水平显著降低（Rhd3、Sod4 和 Ywp1）；（iii）第三类富含碳水化合物活性酶（包括 Cht2、Crh11、Mp65、Pga4、Phr1、Phr2 和 Utr2），但反应有限。据我们所知，这是首次对白色念珠菌菌丝诱导后细胞壁蛋白质组变化进行系统的定量分析。最后，我们提出了新的基于细胞壁蛋白的候选疫苗开发。
