Characterization of Paracoccidioides brasiliensis PbDfg5p, a cell-wall protein implicated in filamentous growth
Characterization of Paracoccidioides brasiliensis PbDfg5p, a cell-wall protein implicated in filamentous growth
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DOI：
10.1002/yea.1574
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发表时间：
2008-02-01
期刊：
影响因子：
2.6
通讯作者：
de Almelda Soares, Celia Maria
中科院分区：
文献类型：
--
作者：
Castro, Nadya Da Silva;Barbosa, Monica Santiago;de Almelda Soares, Celia Maria
巴西副球孢子菌（Paracoccidioides brasiliensis）是拉丁美洲最常见的系统性真菌病的病原体。在人类中，感染开始于吸入真菌繁殖体，其到达肺上皮并分化成酵母寄生阶段。在这里，我们描述了巴西假单胞菌的Dfg 5 p（（d）在bar下有效于（f）在bar下生长（g）在bar下生长）同源物的表征，巴西假单胞菌是一种可预测的细胞壁蛋白，首先在酿酒酵母中鉴定。分析了蛋白质、cDNA和基因组序列。克隆的cDNA在大肠杆菌中表达，纯化的rPbDfg 5 p用于获得多克隆抗体。免疫电子显微镜和生化研究表明，存在的真菌细胞壁中的PbDfg 5 p。酶处理将PbDfg 5 p鉴定为经历N -糖基化的β-葡聚糖连接蛋白。rPbDfg 5 p与细胞外基质组分结合，表明这些相互作用对于导致巴西假单胞菌附着和宿主组织定殖的初始步骤可能是重要的。本文报道的巴西青霉dfg 5基因的核苷酸序列及其推导的蛋白质PbDfg 5 p序列已在GenBank数据库中登录，登录号分别为AY 307855（cDNA）和DQ 534495（genomic）。版权所有（C）2007约翰威利父子有限公司
The dimorphic fungus Paracoccidioides brasiliensis is the causative agent of the most frequent systemic mycosis in Latin America. In humans, infection starts by inhalation of fungal propagules, which reach the pulmonary epithelium and differentiate into the yeast parasitic phase. Here we describe the characterization of a Dfg5p ((d) under bar efective for (f) under bar ilamentous (g) under bar rowth) homologue of P. brasiliensis, a predictable cell wall protein, first identified in Saccharomyces cerevisiae. The protein, the cDNA and genomic sequences were analysed. The cloned cDNA was expressed in Escherichia coli and the purified rPbDfg5p was used to obtain polyclonal antibodies. Immunoelectron microscopy and biochemical studies demonstrated the presence of PbDfg5p in the fungal cell wall. Enzymatic treatments identified PbDfg5p as a beta-glucan linked protein that undergoes N -glycosylation. The rPbDfg5p bound to extracellular matrix components, indicating that those interactions could be important for initial steps leading to P. brasiliensis attachment and colonization of host tissues. The P. brasiliensis dfg5 nucleotide and deduced protein, PbDfg5p, sequences reported in this paper had been submitted to the GenBank database under Accession Nos AY307855 (cDNA) and DQ534495 (genomic). Copyright (C) 2007 John Wiley & Sons, Ltd.