Mycoplasma bovis NADH oxidase functions as both a NADH oxidizing and O 2 reducing enzyme and an adhesin.

Autor: Zhao G; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Zhang H; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Chen X; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Zhu X; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Guo Y; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., He C; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Anwar Khan F; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Chen Y; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China., Hu C; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China., Chen H; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China.; Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, China.; Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, Huazhong Agricultural University, Wuhan, 430070, China., Guo A; National Key Laboratory of Agricultural Microbiology, Huazhong Agriculture University, Wuhan, 430070, China. aizhen@mail.hzau.edu.cn.; College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China. aizhen@mail.hzau.edu.cn.; Key Laboratory of Development of Veterinary Diagnostic Products, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, 430070, China. aizhen@mail.hzau.edu.cn.; Hubei International Scientific and Technological Cooperation Base of Veterinary Epidemiology, Huazhong Agricultural University, Wuhan, 430070, China. aizhen@mail.hzau.edu.cn.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2017 Mar 03; Vol. 7 (1), pp. 44. Date of Electronic Publication: 2017 Mar 03.
DOI: 10.1038/s41598-017-00121-y
Abstrakt: Mycoplasma bovis causes considerable economic losses in the cattle industry worldwide. In mycoplasmal infections, adhesion to the host cell is of the utmost importance. In this study, the amino acid sequence of NOX was predicted to have enzymatic domains. The nox gene was then cloned and expressed in Escherichia coli. The enzymatic activity of recombinant NOX (rNOX) was confirmed based on its capacity to oxidize NADH to NAD + and reduce O 2 to H 2 O 2 . The adherence of rNOX to embryonic bovine lung (EBL) cells was confirmed with confocal laser scanning microscopy, enzyme-linked immunosorbent assay, and flow cytometry. Both preblocking EBL cells with purified rNOX and preneutralizing M. bovis with polyclonal antiserum to rNOX significantly reduced the adherence of M. bovis to EBL cells. Mycoplasma bovis NOX- expressed a truncated NOX protein at a level 10-fold less than that of the wild type. The capacities of M. bovis NOX- for cell adhesion and H 2 O 2 production were also significantly reduced. The rNOX was further used to pan phage displaying lung cDNA library and fibronectin was determined to be potential ligand. In conclusion, M. bovis NOX functions as both an active NADH oxidase and adhesin, and is therefore a potential virulence factor.
Databáze: MEDLINE
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