The mpn668 gene of Mycoplasma pneumoniae encodes a novel organic hydroperoxide resistance protein.

Mycoplasma pneumoniae (M. pneumoniae), as an obligate parasite, has evolved a protective strategy for coping with oxidative challenges caused by M. pneumoniae itself as well as the host immune system. However, to date, few antioxidant enzymes have been identified in mycoplasmas. In this report, we identified a protein encoded by the mpn668 gene from M. pneumoniae with a putative function as an organic hydroperoxide reductase (Ohr). The results indicated that the recombinant 140 amino acid protein, designated rMPN668, displayed hydroperoxidase activity towards both organic (tert-butyl hydroperoxide) and inorganic (hydrogen peroxide) hydroperoxides in the presence of a reducing agent such as dithiothreitol. Moreover, the expression of mpn668 in M. pneumoniae is upregulated in response to oxidative stress. Additionally, homology modeling of MPN668 and a molecular dynamics simulation suggest that both Cys55 and Cys119 form part of the active site of the protein. Mutants in which Cys55 or Cys119 were replaced with a serine lack antioxidant activity, indicating that MPN668 is a Cys-based peroxidase, consistent with it representing a new member of the Ohr family.