Mycoplasma bovis NADH oxidase functions as both a NADH oxidizing and O2 reducing enzyme and an adhesin
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 i...
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Published in | Scientific reports Vol. 7; no. 1; p. 44 |
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Main Authors | , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
London
Nature Publishing Group UK
03.03.2017
Nature Portfolio |
Subjects | |
Online Access | Get full text |
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Summary: | 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. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 2045-2322 2045-2322 |
DOI: | 10.1038/s41598-017-00121-y |