Thevissen Karin, François Isabelle E J A, Takemoto Jon Y, Ferket Kathelijne K A, Meert Els M K, Cammue Bruno P A
CMPG (Centre of Microbial and Plant Genetics), Katholieke Universiteit Leuven, Kasteelpark Arenberg 20, B-3001, Heverlee, Belgium.
FEMS Microbiol Lett. 2003 Sep 12;226(1):169-73. doi: 10.1016/S0378-1097(03)00590-1.
DmAMP1, an antifungal plant defensin from Dahlia merckii, was shown previously to require the presence of sphingolipids for fungicidal action against Saccharomyces cerevisiae. Sphingolipids may stabilize glycosylphosphatidylinositol (GPI)-anchored proteins, which interact with DmAMP1, or they may directly serve as DmAMP1 binding sites. In the present study, we demonstrate that S. cerevisiae disruptants in GPI-anchored proteins showed small or no increased resistance towards DmAMP1 indicating no involvement of these proteins in DmAMP1 action. Further, studies using an enzyme-linked immunosorbent assay (ELISA)-based binding assay revealed that DmAMP1 interacts directly with sphingolipids isolated from S. cerevisiae and that this interaction is enhanced in the presence of equimolar concentrations of ergosterol. Therefore, DmAMP1 antifungal action involving membrane interaction with sphingolipids and ergosterol is proposed.
DmAMP1是一种来自大丽花的抗真菌植物防御素,先前已证明它对酿酒酵母的杀真菌作用需要鞘脂的存在。鞘脂可能会稳定与DmAMP1相互作用的糖基磷脂酰肌醇(GPI)锚定蛋白,或者它们可能直接作为DmAMP1的结合位点。在本研究中,我们证明GPI锚定蛋白中的酿酒酵母破坏株对DmAMP1的抗性增加很小或没有增加,表明这些蛋白不参与DmAMP1的作用。此外,使用基于酶联免疫吸附测定(ELISA)的结合试验的研究表明,DmAMP1与从酿酒酵母中分离的鞘脂直接相互作用,并且在等摩尔浓度的麦角固醇存在下这种相互作用会增强。因此,提出了DmAMP1的抗真菌作用涉及与鞘脂和麦角固醇的膜相互作用。