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Fructose-1,6-bisphosphate aldolase is involved in Mycoplasma bovis colonization as a fibronectin-binding adhesin

Huang, Jing, Zhu, Hongmei, Wang, Jiayao, Guo, Yongpeng, Zhi, Ye, Wei, Haohua, Li, Hanxiong, Guo, Aizhen, Liu, Dongming, Chen, Xi
Research in veterinary science 2019 v.124 pp. 70-78
Mycoplasma bovis, Western blotting, adhesins, adhesion, adhesion inhibition assay, bacterial colonization, cattle, cell adhesion, cytoplasm, dose response, enzyme activity, enzyme-linked immunosorbent assay, fibronectins, fluorescent antibody technique, fructose, fructose-bisphosphate aldolase, genes, host-cell factors, lungs, pathogens, polyclonal antibodies, rabbits, recombinant proteins, virulence
Mycoplasma bovis is a common pathogenic microorganism of cattle and represents an important hazard on the cattle industry. Adherence to host cells is a significant component of mycoplasma-pathogenesis research. Fibronectin (Fn), an extracellular matrix protein, is a common host cell factor that can interact with the adhesions of pathogens. The aims of this study were to investigate the Fn-binding properties of M. bovis fructose-1,6-bisphosphate aldolase (FBA) and evaluate its role as a cell adhesion factor during mycoplasma colonization. The fba (MBOV_RS00435) gene of M. bovis was cloned and expressed, with the resulting recombinant protein used to prepare rabbit polyclonal antibodies. The purified recombinant FBA (rFBA) was shown to have fructose bisphosphate aldolase activity. Western blot indicated that FBA was an antigenically conserved protein in several M. bovis strains. Western blot combined with immunofluorescent assay (IFA) revealed that FBA was dual-localized to both cytoplasm and membrane in M. bovis. IFA showed that rFBA was able to adhere to embryonic bovine lung (EBL) cells. Meanwhile, an adhesion inhibition assay demonstrated that anti-rFBA antibodies could significantly block the adhesion of M. bovis to EBL cells. Moreover, a dose-dependent binding of rFBA to Fn was found by dot blotting and enzyme-linked immunosorbent assays. Together these results provided evidence that FBA is a surface-localized and antigenic protein of M. bovis, suggesting that it may function as a virulence determinant through interacting with host Fn.