关键词: 双向电泳, 蛋白, 糠秕马拉色菌

Abstract: Objectives To identify differential proteins in the yeast and mycelial phases of Malassezia furfur by using the two-dimensional electrophoresis (2-DE) protocol and tandem spectrum (MS/MS), and to understand the pathogenic mechanism of M.furfur and its morphological switching mechanism at protein level as well.Methods The type strain of M.furfur was inoculated in the yeast and mycelial phase media respectively. Vortexing with glass beads and ultrasonication were used to break up the cell wall into pieces, and trichloroacetate/acetone precipitation was applied to obtain the total proteins. These proteins were separated and visualized by 2-DE, analyzed by PDQuest soft to detect the differentially expressed protein spots. Electrospray-tandem spectrum analysis combined with homology search by Mascot and NCBI's Blast was used to identify proteins.Results The stable protein profiles from the mycelial and yeast phases were compared, more than 800 spots were detected respectively. Of these, 64 spots were regulated more than 3-fold, 11 were identified only in the yeast phases and 9 were identified only in the mycelial phases. Selected 40 differential spots, 22 were identified, corresponding to 16 unique proteins. In the identified proteins,9 proteins including actin, cofilin etc. were up-regulated and 5 proteins including glutathione transferase, cytoskeletal signaling protein etc, were down-regulated in mycelial phase.Conclusions Sixteen protein spots implicated in some cellular processes, such as metobalism, motility, oxidase stress etc. were positively identified by experiencements and important information would be provided for understanding the mechanisms underlying phenotype transformation of M.furfur and its pathogenesis.

Key words: two-dimensional electrophoresis, protein, Malassezia furfur