The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering.
The N-linking glycosylation system from Actinobacillus pleuropneumoniae is required for adhesion and has potential use in glycoengineering.
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DOI：
10.1098/rsob.160212
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发表时间：
2017-01
期刊：
影响因子：
5.8
通讯作者：
BRaDP1T Consortium
中科院分区：
文献类型：
--
作者：
Cuccui J;Terra VS;Bossé JT;Naegeli A;Abouelhadid S;Li Y;Lin CW;Vohra P;Tucker AW;Rycroft AN;Maskell DJ;Aebi M;Langford PR;Wren BW;BRaDP1T Consortium
胸膜肺炎放线杆菌是引起猪传染性胸膜肺炎的一种粘膜呼吸道病原菌。发病机制的研究表明，胶囊，外毒素和外膜蛋白的主要作用。胸膜肺炎放线杆菌也可以糖基化蛋白质，使用细胞质N-连接的糖基化酶命名为NGT，但其转录排列和毒力的作用仍然未知。我们研究了NGT位点，并证明了推定的转录单位由rimO，NGT和一个糖基转移酶称为AGT。根据这些信息，我们使用了A。在大肠杆菌内，使用与抗葡聚糖抗体反应的己糖聚合物修饰胸膜肺炎糖基化基因座的受体蛋白。对截短蛋白质的质谱分析显示，该操纵子可以将多达29个重复单元添加到适当的序列子。我们通过创建缺失突变体并在一种新型呼吸道细胞系粘附模型中对其进行测试，证明了NGT在毒力中的重要性。这项研究表明NGT糖基化系统的发病机制和糖工程的潜在生物技术应用的重要性。
Actinobacillus pleuropneumoniae is a mucosal respiratory pathogen causing contagious porcine pleuropneumonia. Pathogenesis studies have demonstrated a major role for the capsule, exotoxins and outer membrane proteins. Actinobacillus pleuropneumoniae can also glycosylate proteins, using a cytoplasmic N-linked glycosylating enzyme designated NGT, but its transcriptional arrangement and role in virulence remains unknown. We investigated the NGT locus and demonstrated that the putative transcriptional unit consists of rimO, ngt and a glycosyltransferase termed agt. From this information we used the A. pleuropneumoniae glycosylation locus to decorate an acceptor protein, within Escherichia coli, with a hexose polymer that reacted with an anti-dextran antibody. Mass spectrometry analysis of a truncated protein revealed that this operon could add up to 29 repeat units to the appropriate sequon. We demonstrated the importance of NGT in virulence, by creating deletion mutants and testing them in a novel respiratory cell line adhesion model. This study demonstrates the importance of the NGT glycosylation system for pathogenesis and its potential biotechnological application for glycoengineering.