Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, 163319, China.
Heilongjiang Provincial Key Laboratory of Prevention and Control of Bovine Diseases, College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing, 163319, China.
Anaerobe. 2020 Jun;63:102184. doi: 10.1016/j.anaerobe.2020.102184. Epub 2020 Mar 4.
Fusobacterium necrophorum is a Gram negative, spore-free, anaerobic bacterium that can cause pyogenic and necrotic infections in animals and humans. It is a major bovine pathogen and causes hepatic abscesses, foot rot, and necrotic laryngitis. The 43K OMP of F. necrophorum is an outer membrane protein with molecular weight of 43 kDa, exhibiting similarity to pore-forming proteins of other Fusobacterium species that plays an important role in bacterial infections. However, the role of 43K OMP in F. necrophorum adhesion remains unknown. In this study, we evaluated whether the 43K OMP of F. necrophorum mediates adhesion to BHK-21 cells and performed a preliminary screen of the proteins that interact with 43K OMP of F. necrophorum by immunoprecipitation-mass spectrometry. The results showed that the natural 43K OMP and recombinant 43K OMP could bind to BHK-21 cells, and preincubation of F. necrophorum with an antibody against the recombinant 43K OMP of F. necrophorum decreased binding to BHK-21 cells. Seventy differential interacting proteins were successfully screened by immunoprecipitation-mass spectrometry. Among these seventy differential interacting proteins, seven cell membrane proteins and four extracellular matrix proteins shown to be relevant to bacteria adhesion through subcellular localization and single-molecule function analysis. These data increase our understanding of the pathogenesis of F. necrophorum and provide a new theoretical basis for the design of antimicrobial drugs against F. necrophorum.
坏死梭杆菌是一种革兰氏阴性、无孢子、厌氧细菌，可导致动物和人类化脓性和坏死性感染。它是一种主要的牛病原体，可引起肝脓肿、腐蹄病和坏死性喉炎。坏死梭杆菌的 43K OMP 是一种外膜蛋白，分子量为 43 kDa，与其他梭杆菌属的孔形成蛋白具有相似性，在细菌感染中发挥重要作用。然而，43K OMP 在坏死梭杆菌黏附中的作用尚不清楚。在本研究中，我们评估了坏死梭杆菌的 43K OMP 是否介导对 BHK-21 细胞的黏附，并通过免疫沉淀-质谱法对与坏死梭杆菌的 43K OMP 相互作用的蛋白质进行了初步筛选。结果表明，天然 43K OMP 和重组 43K OMP 均可与 BHK-21 细胞结合，用针对坏死梭杆菌的重组 43K OMP 的抗体预先孵育坏死梭杆菌可降低与 BHK-21 细胞的结合。通过免疫沉淀-质谱法成功筛选出 70 种差异相互作用蛋白。在这 70 种差异相互作用蛋白中，有 7 种细胞膜蛋白和 4 种细胞外基质蛋白通过亚细胞定位和单分子功能分析显示与细菌黏附有关。这些数据增加了我们对坏死梭杆菌发病机制的理解，并为设计针对坏死梭杆菌的抗菌药物提供了新的理论依据。
