The antifungal properties of a 2S albumin-homologous protein from passion fruit seeds involve plasma membrane permeabilization and ultrastructural alterations in yeast cells

被引:37
作者
Agizzio, Ana Paula
Da Cunha, Maura
Carvalho, Andre O.
Oliveira, Marco Antonio
Ribeiro, Suzanna F. F.
Gomes, Valdirene M. [1 ]
机构
[1] Univ Estadual Norte Fluminense, Ctr Biociencias & Biotecnol, Lab Fisiol & Bioquim Microorganismos, BR-28013602 Campos Dos Goytacazes, RJ, Brazil
[2] Univ Estadual Norte Fluminense, Ctr Biociencias & Biotecnol, Lab Biol Celular & Tecidual, BR-28013602 Campos Dos Goytacazes, RJ, Brazil
[3] Univ Vale Paraiba, Inst Pesquisa & Desenvolvimento, San Jose Dos Campus, SP, Brazil
关键词
2S albumins; antifungal protein; fungal cell wall; membrane permeabilization; Saccharomyces cerevisiae;
D O I
10.1016/j.plantsci.2006.06.001
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Different types of antimicrobial proteins were purified from plant seeds, including chitinases, beta-1,3-glucanases, defensins, thionins, lipid transfer proteins and 2S albumins. It has become clear that these groups of proteins play an important role in the protection of plants from microbial infection. Recent results from our laboratory have shown that the defense-related proteins from passion fruit seeds, named Pf1 and Pf2 (which show sequence homology with 2S albumins), inhibit fungal growth and glucose-stimulated acidification of the medium by Saccharomyces cerevisiae cells. The aim of this study was to determine whether 2S albumins from passion fruit seeds induce plasma membrane permeabilization and cause morphological alterations in yeast cells. Initially, we used an assay based on the uptake of SYTOX Green, an organic compound that fluoresces upon interaction with nucleic acids and penetrates cells with compromised plasma membranes, to investigate membrane permeabilization in S. cerevisiae cells. When viewed with a confocal laser microscope, S. cervisiae cells showed strong SYTOX Green fluorescence in the cytosol, especially in the nuclei. 2S albumins also inhibited glucose-stimulated acidification of the medium by S. cerevisiae cells, which indicates a probable impairment of fungal metabolism. The microscopical analysis of the yeast cells treated with 2S albumins demonstrated several morphological alterations in cell shape, cell surface, cell wall and bud formation, as well as in the organization of intracellular organelles. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
引用
收藏
页码:515 / 522
页数:8
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