The study of differentially expressed proteins in yeast and mycelial phases of Malassiza furfur

XIANG Yun; RAN Yu-ping; TONG Ai-ping; GOU Lan-tu; WANG Wei; DAI Ya-ling

doi:10.3969/j.issn.1673-3827.2014.05.001

Abstract：Objectives To identify differential proteins in the yeast and mycelial phases of Malassezia furfur by using the two-di-mensional 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 vis-ualized by 2-DE,analyzed by PDQuest soft to detect the differentially expressed protein spots Electrospray-tandem spectrum analy-sis combined with homology search by Mascot and NCBIˊs 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 regula-ted more than 3-fold,11 were identified only in the yeast phases and 9 were identified only in the mycelial phases Selected 40 dif-ferential 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 mecha-nisms underlying phenotype transformation of M. furfur and its pathogenesis.