Molecular and immunological characterization of Pasteurella multocida serotype A:3 OmpA:: evidence of its role in P-multocida interaction with extracellular matrix molecules
Molecular and immunological characterization of Pasteurella multocida serotype A:3 OmpA:: evidence of its role in P-multocida interaction with extracellular matrix molecules
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DOI：
10.1016/s0882-4010(03)00098-6
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发表时间：
2003-10-01
影响因子：
3.8
通讯作者：
Quijano-Blas, RA
中科院分区：
文献类型：
--
作者：
Dabo, SM;Confer, AW;Quijano-Blas, RA
克隆并鉴定了多杀性巴氏杆菌ompa样基因（PmOmpA）。成熟蛋白分子量为35,075 Da，与大肠杆菌OmpA蛋白具有显著的相似性。膜拓扑分析预测，与大肠杆菌OmpA一样，PmOmpA的n端一半以八链跨膜反平行β -桶的形式存在，显示出四个可变的亲水性和表面暴露区域，预测抗原峰可能涉及血清抵抗或粘附。此外，在PmOmpA中，n端p桶和c端外质结构域之间的Ala-Pro重复区完全缺失。PmOmpA在大肠杆菌中表达，免疫印迹分析显示重组PmOmpA具有免疫原性，并在体内表达。证实了PmOmpA与生物素化的马丁达比牛肾（MDBK）细胞表面蛋白、纤维连接蛋白和肝素的结合。此外，PmOmpA与MDBK单层结合，用抗PmOmpA预处理多杀假单胞菌全细胞可显著降低对纤维连接蛋白的粘附。配体印迹分析显示，分别在37℃和100℃加热时，纤维连接蛋白与PmOmpA的天然形式和热修饰形式结合。总的来说，这些数据表明PmOmpA可能通过肝素和/或纤维连接蛋白桥接参与了多毒杆菌232对宿主细胞的粘附。(C) 2003 Elsevier Ltd.版权所有。
Pasteurella multocida OmpA-like gene (PmOmpA) was cloned and characterized. The mature protein had a molecular mass of 35,075 Da and significant similarity with Escherichia coli (E. coli) OmpA proteins. Membrane topology analyses predict that like E. coli OmpA, the N-terminal half of PmOmpA exists as an eight-stranded transmembrane antiparallel beta-barrel that displays four variable hydrophilic and surface-exposed regions with predicted antigenic peaks that may be involved in serum resistance or adherence. In addition, the Ala-Pro repeat region between the N-terminal P-barrel and C-terminal periplasmic domains is completely missing in PmOmpA. PmOmpA was expressed in E. coli and immunoblots analysis revealed that the recombinant PmOmpA was immunogenic, and expressed in vivo. The binding of PmOmpA to biotinylated Madin Darby bovine kidney (MDBK) cells surface proteins, fibronectin and heparin was demonstrated. Furthermore, PmOmpA binds MDBK monolayers and pre-treatment of P. multocida whole cells with anti-PmOmpA significantly reduced adherence to fibronectin. Ligand blot analysis revealed that fibronectin binds to the native and heat modified forms of PmOmpA when heated at 37 and 100 degreesC, respectively. Collectively these data indicate that PmOmpA may be involved in P. multocida 232 adherence to host cells via heparin and/or fibronectin bridging. (C) 2003 Elsevier Ltd. All rights reserved.