Identification and characterization of a novel stress-responsive outer membrane protein Lip40 from Actinobacillus pleuropneumoniae.
Identification and characterization of a novel stress-responsive outer membrane protein Lip40 from Actinobacillus pleuropneumoniae.
复制标题
来自静脉肌杆菌的新型应力反应性外膜蛋白LIP40的鉴定和表征。
DOI：
10.1186/s12896-015-0199-8
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发表时间：
2015-11-25
影响因子：
3.5
通讯作者：
Liu J
中科院分区：
文献类型：
--
作者：
Hu X;Yan H;Liu K;Hu J;Qi C;Yang J;Liu Y;Zhao J;Liu J
胸膜肺炎放线杆菌是一种革兰氏阴性细菌，是猪胸膜肺炎的病原体，猪胸膜肺炎是一种高度传染性且常常致命的疾病。由于目前的疫苗对胸膜肺炎放线菌感染的保护有限，因此迫切需要开发更有效的疫苗。免疫原性和保护性抗原（例如外膜脂蛋白）的鉴定将促进这一目的。使用多种算法从胸膜肺炎放线菌的基因组序列中预测出 60 种假定的脂蛋白。在这里，我们重点关注来自胸膜肺炎放线菌菌株 SLW01（血清型 1）的推定脂蛋白 Lip40 的特征。 Lip40与许多细菌脂蛋白具有序列相似性，并且Lip40的结构预测表明它与胸膜肺炎放线菌TbpB相似。 Lip40 的 N 末端包含一个有趣的串联重复序列 Q(E/D/P)QPK。实时 RT-PCR 表明 lip40 的表达在 42°C、16°C 和厌氧条件下显着上调。大肠杆菌 BL21(DE3) 中产生的重组 Lip40 (rLip40) 可被针对胸膜肺炎放线菌的猪恢复期血清特异性识别。 Lip40被证实定位于细菌外膜，并且当胸膜肺炎放线杆菌在各种应激条件下培养时，其表达显着增加。 Lip40 还保护 75% 的小鼠免受致命的致命性胸膜肺炎放线菌感染。免疫原性外膜蛋白 Lip40 具有应激反应性，可以保护小鼠免受感染，并且可能是毒力决定因素。对 Lip40 的进一步研究应该会加快疫苗的开发，并深入了解胸膜肺炎放线菌的发病机制。本文的在线版本 (doi:10.1186/s12896-015-0199-8) 包含补充材料，可供授权用户使用。
Actinobacillus pleuropneumoniae, a Gram-negative bacterium, is the causative agent of porcine pleuropneumonia, a highly contagious and often fatal disease. Because current vaccines confer limited protection against A. pleuropneumoniae infection, the development of more effective vaccines is urgently required. The identification of immunogenic and protective antigens, such as an outer-membrane lipoprotein, will advance this purpose. Sixty putative lipoproteins were predicted from the genomic sequence of A. pleuropneumoniae using multiple algorithms. Here, we focused on the characteristics of the putative lipoprotein Lip40 from A. pleuropneumoniae strain SLW01 (serovar 1). Lip40 shares sequence similarity with many bacterial lipoproteins, and the structural prediction of Lip40 suggests that it is similar to A. pleuropneumoniae TbpB. The N-terminus of Lip40 contains an interesting tandemly repeated sequence, Q(E/D/P)QPK. Real-time RT–PCR indicated that the expression of lip40 was significantly upregulated at 42 °C, at 16 °C, and under anaerobic conditions. Recombinant Lip40 (rLip40) produced in Escherichia coli BL21(DE3) was specifically recognized by porcine convalescent serum directed against A. pleuropneumoniae. Lip40 was confirmed to localize at the bacterial outer membrane, and its expression was significantly stimulated when A. pleuropneumoniae was cultured under various stress conditions. Lip40 also protected 75 % of mice from fatal virulent A. pleuropneumoniae infection. The immunogenic outer-membrane protein Lip40 is stress responsive, protects mice against infection, and might be a virulence determinant. Further investigation of Lip40 should expedite vaccine development and provide insight into the pathogenesis of A. pleuropneumoniae. The online version of this article (doi:10.1186/s12896-015-0199-8) contains supplementary material, which is available to authorized users.