Arioka M, Hirata A, Takatsuki A, Yamasaki M
Department of Agricultural Chemistry, University of Tokyo, Japan.
J Gen Microbiol. 1991 Jun;137(6):1253-62. doi: 10.1099/00221287-137-6-1253.
Brefeldin A (BFA) inhibited in a dose-dependent manner secretion of the cell-surface enzyme acid phosphatase (APase) into the periplasm of Candida albicans and caused intracellular accumulation of enzyme protein. Cells grown in the presence of BFA became more dense, implying that cell-surface growth was also blocked by BFA treatment. The APase that was accumulated intracellularly migrated faster on SDS-PAGE, suggesting less N-linked glycosylation compared with the mature, periplasmic APase produced in the absence of BFA. Pulse-chase experiments and gel-filtration of oligosaccharides released by Endo H treatment suggested that the core-glycosylated precursor form of APase accumulated in the presence of BFA. These results strongly suggested that endoplasmic reticulum (ER)-to-Golgi transport in C. albicans was inhibited by BFA. Aberrant membrane structures were observed in BFA-treated cells. Within 1 h of BFA removal these structures were replaced with rough ER membranes, suggesting that the accumulated membranes were derived from the ER.
布雷菲德菌素A（BFA）以剂量依赖性方式抑制白色念珠菌细胞表面酶酸性磷酸酶（APase）向周质的分泌，并导致酶蛋白在细胞内积累。在BFA存在下生长的细胞变得更加致密，这意味着细胞表面生长也被BFA处理所阻断。细胞内积累的APase在SDS-PAGE上迁移得更快，这表明与在没有BFA的情况下产生的成熟周质APase相比，其N-连接糖基化程度更低。脉冲追踪实验和内切糖苷酶H处理释放的寡糖的凝胶过滤表明，在BFA存在下积累了APase的核心糖基化前体形式。这些结果强烈表明，BFA抑制了白色念珠菌内质网（ER）到高尔基体的转运。在BFA处理的细胞中观察到异常的膜结构。在去除BFA后1小时内，这些结构被粗糙内质网的膜所取代，这表明积累的膜来源于内质网。
