title: Respiratory detoxification of nitric oxide by the cytochrome c nitrite reductase of Escherichia coli.
abstract: Nitric oxide is a key element in host defense against invasive pathogens. The periplasmic cytochrome c nitrite reductase (NrfA) of Escherichia coli catalyzes the respiratory reduction of nitrite, but in vitro studies have shown that it can also reduce nitric oxide. The physiological significance of the latter reaction in vivo has never been assessed. In this study the reduction of nitric oxide by Escherichia coli was measured in strains active or deficient in periplasmic nitrite reduction. Nrf(+) cells, harvested from cultures grown anaerobically, possessed a nitric-oxide reductase activity with physiological electron donation of 60 nmol min(-1) x mg dry wt(-1), and an in vivo turnover number of NrfA of 390 NO* s(-1) was calculated. Nitric-oxide reductase activity could not be detected in Nrf(-) strains. Comparison of the anaerobic growth of Nrf(+) and Nrf(-) strains revealed a higher sensitivity to nitric oxide in the NrfA(-) strains. A higher sensitivity to the nitrosating agent S-nitroso-N-acetyl penicillamine (SNAP) was also observed in agar plate disk-diffusion assays. Oxygen respiration by E. coli was also more sensitive to nitric oxide in the Nrf(-) strains compared with the Nrf(+) parent strain. The results demonstrate that active periplasmic cytochrome c nitrite reductase can confer the capacity for nitric oxide reduction and detoxification on E. coli. Genomic analysis of many pathogenic enteric bacteria reveals the presence of nrf genes. The present study raises the possibility that this reflects an important role for the cytochrome c nitrite reductase in nitric oxide management in oxygen-limited environments.
authors.affiliation: Centre for Metalloprotein Spectroscopy and Biology, School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, United Kingdom.
authors.forename: Susannah R
authors.initials: SR
authors.lastname: Poock
authors.forename: Emily R
authors.initials: ER
authors.lastname: Leach
authors.forename: James W B
authors.initials: JW
authors.lastname: Moir
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Cole
authors.forename: David J
authors.initials: DJ
authors.lastname: Richardson
mesh_terms: D000693:Anaerobiosis; D045263:Cytochromes a1; D003575:Cytochromes c1; D004926:Escherichia coli / Q000378:metabolism*; D009565:Nitrate Reductases / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction; D010101:Oxygen Consumption; D010396:Penicillamine / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: C110881:S-nitro-N-acetylpenicillamine; D009569:Nitric Oxide; D045263:Cytochromes a1; D003575:Cytochromes c1; D009565:Nitrate Reductases; C054690:nitrate reductase (cytochrome); D010396:Penicillamine