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鉴定一种外膜脂蛋白,该脂蛋白参与了在动物模型中卡他莫拉菌的鼻咽定植。

Identification of an outer membrane lipoprotein involved in nasopharyngeal colonization by Moraxella catarrhalis in an animal model.

机构信息

Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, Maryland, USA.

出版信息

Infect Immun. 2014 Jun;82(6):2287-99. doi: 10.1128/IAI.01745-14. Epub 2014 Mar 18.

Abstract

Colonization of the human nasopharynx by Moraxella catarrhalis is presumed to involve attachment of this bacterium to the mucosa. DNA microarray analysis was used to determine whether attachment of M. catarrhalis to human bronchial epithelial (HBE) cells in vitro affected gene expression in this bacterium. Attachment affected expression of at least 454 different genes, with 163 being upregulated and 291 being downregulated. Among the upregulated genes was one (ORF113) previously annotated as encoding a protein with some similarity to outer membrane protein A (OmpA). The protein encoded by ORF113 was predicted to have a signal peptidase II cleavage site, and globomycin inhibition experiments confirmed that this protein was indeed a lipoprotein. The ORF113 protein also contained a predicted peptidoglycan-binding domain in its C-terminal half. The use of mutant and recombinant M. catarrhalis strains confirmed that the ORF113 protein was present in outer membrane preparations, and this protein was also shown to be at least partially exposed on the bacterial cell surface. A mutant unable to produce the ORF113 protein showed little or no change in its growth rate in vitro, in its ability to attach to HBE cells in vitro, or in its autoagglutination characteristics, but it did exhibit a reduced ability to survive in the chinchilla nasopharynx. This is the first report of a lipoprotein essential to the ability of M. catarrhalis to persist in an animal model.

摘要

人鼻咽部的莫拉氏菌定植被认为涉及到这种细菌与黏膜的附着。DNA 微阵列分析被用来确定莫拉氏菌在体外附着于人体支气管上皮 (HBE) 细胞是否会影响该细菌的基因表达。附着影响了至少 454 个不同基因的表达,其中 163 个上调,291 个下调。上调的基因之一 (ORF113) 之前被注释为编码一种与外膜蛋白 A (OmpA) 具有一定相似性的蛋白质。ORF113 编码的蛋白质被预测具有信号肽酶 II 切割位点,并且 globomycin 抑制实验证实该蛋白质确实是一种脂蛋白。ORF113 蛋白在其 C 端的一半还包含一个预测的肽聚糖结合结构域。使用突变体和重组莫拉氏菌菌株证实,ORF113 蛋白存在于外膜制剂中,并且该蛋白至少部分暴露在细菌细胞表面。一个不能产生 ORF113 蛋白的突变体在体外生长速度、在体外附着于 HBE 细胞的能力或自身聚集特性方面几乎没有变化,但它确实表现出在豚鼠鼻咽部生存能力降低的现象。这是首次报道脂蛋白对莫拉氏菌在动物模型中持续存在的能力至关重要。

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