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The Antifungal Activity of RsAFP2, a Plant Defensin from Raphanus sativus, Involves the Induction of Reactive Oxygen Species in Candida albicans

The Antifungal Activity of RsAFP2, a Plant Defensin from Raphanus sativus, Involves the Induction of Reactive Oxygen Species in Candida albicans,10.11

The Antifungal Activity of RsAFP2, a Plant Defensin from Raphanus sativus, Involves the Induction of Reactive Oxygen Species in Candida albicans   (Citations: 16)
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RsAFP2 (Raphanus sativus antifungal peptide 2), an antifungal plant defensin isolated from seed of R. sativus, interacts with glucosylceramides (GlcCer) in membranes of susceptible yeast and fungi and induces membrane permeabilization and fungal cell death. However, using carboxyfluorescein-containing small unilamellar vesicles containing purified GlcCer, we could not observe permeabilization as a consequence of insertion of RsAFP2 in such vesicles. Therefore, we focused on a putative RsAFP2-induced signaling cascade downstream of RsAFP2-binding to GlcCer in fungal membranes. We show that RsAFP2 induces reactive oxygen species (ROS) in Candida albicans wild type in a dose-dependent manner, but not at all in an RsAFP2-resistant ΔgcsC. albicans mutant that lacks the RsAFP2-binding site in its membranes. These findings indicate that upstream binding of RsAFP2 to GlcCer is needed for ROS production leading to yeast cell death. Moreover, the antioxidant ascorbic acid blocks RsAFP2-induced ROS generation, as well as RsAFP2 antifungal activity. These data point to the presence of an intracellular plant defensin-induced signaling cascade, which involves ROS generation and leads to fungal cell growth arrest.
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    • ...Moreover, the antioxidant ascorbic acid blocks Rs-AFP2-induced ROS generation and Rs-AFP2 antifungal activity [1]...
    • ...It has been demonstrated that some plant defensins could induce oxidative stress in fungal cell exposed to them [1]...
    • ...difluorofluorescein diacetate (Calbiochem - EMD), respectively, using methods described by [1] with some modifications...
    • ...Different defensins have also been found to permeabilize membranes and to modulate ion flux across membranes, and they have since become popular models for understanding how ion channel proteins function [1]...
    • ...Lobo et al. [10] have demonstrated that PsD1 interacts with cyclin F and consequently interferes with the cell cycle [1, 27]; NaD1 and Rs-AFP2 induce ROS production; and that these DIC Fluorescence...

    Erica O. Melloet al. Antifungal Activity of Pv D1 Defensin Involves Plasma Membrane Permeab...

    • ...It interacts with glucosylceramides in fungal and yeast membranes, induces membrane permeabilization and production of reactive oxygen species, resulting in cessation of growth and cell death (Aerts et al. 2007)...

    Jack H. Wonget al. Proteins with antifungal properties and other medicinal applications f...

    • ...Interestingly, in Candida albicans , the anti-fungal activity of one plant defensin RsAFP2, which binds glucosylceramide, was found to involve the production of reactive oxygen species (ROS) suggesting that binding to the membrane ligand initiated a signal transduction cascade that culminated in the production of ROS and membrane permeabilisation (Fig. 3b ) (Aerts et al., 2007 ; Thevissen et al., 2007) . Despite the progress that has been ...

    John P. Morrissey. Biological Activity of Defence-Related Plant Secondary Metabolites

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