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.

坏死镰刀菌是革兰氏阴性，无孢子的厌氧细菌，可引起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，足腐烂和坏死性喉炎。的43K OMP F. necrophorum是外膜蛋白与分子量43千道尔顿，表现出相似性的其他成孔蛋白梭杆菌播放在细菌感染中起重要作用的物种。但是，尚不清楚43K OMP在坏死镰刀菌粘附中的作用。在这项研究中，我们评估了F. necrophorum的43K OMP是否介导了对BHK-21细胞的粘附，并对与KHK -21的43K OMP相互作用的蛋白质进行了初步筛选。通过免疫沉淀质谱法测定死灵藻。结果表明，天然43K OMP和重组43K OMP可以结合BHK-21细胞，并预温育F. necrophorum与针对重组43K OMP的抗体F. necrophorum降低结合至BHK-21细胞。通过免疫沉淀-质谱法成功筛选了七十种不同的相互作用蛋白。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粘附有关。这些数据增加了我们对坏死镰刀菌发病机理的了解。为抗坏死镰刀菌的抗菌药物设计提供新的理论依据。