title: Determinants of nitric oxide steady-state levels during anaerobic respiration by Neisseria gonorrhoeae.
abstract: Nitric oxide (NO) is an important host defence molecule that varies its immune stimulatory effects depending on the concentrations at which it is produced, with low concentrations (< 1 microM) promoting an anti-inflammatory host response while higher concentrations (>1 microM) lead to inflammatory responses. Neisseria gonorrhoeae grows anaerobically by anaerobic respiration using nitrite reductase (Nir) to convert nitrite to NO and nitric oxide reductase (Nor) to convert NO to nitrous oxide. As N. gonorrhoeae can both produce and degrade NO, we have begun a study of NO metabolism in this bacterium to understand how gonococcal manipulation of NO concentration may influence the inflammatory response during infection. N. gonorrhoeae has an apparent Nir Km of 33 microM nitrite and an apparent Nor Km of 1.2 microM NO. The maximum specific activities for Nir and Nor were 135 nmoles nitrite reduced per minute per OD600 (pH 6.7) and 270 nmoles NO reduced per minute per OD600 (pH 7.5) respectively. N. gonorrhoeae established a steady-state concentration of NO after nitrite addition that was dependent on the nitrite concentration until saturation at 1 mM nitrite. The NO steady-state level decreased as pH increased, and the ratio of activities of Nir and Nor correlated to the NO steady-state level. When the NO donor DETA/NO was used to simulate host NO production, N. gonorrhoeae also established a NO steady-state level. The concentration of NO at steady state was found to be a function of the concentration of NO generated by DETA/NO, with N. gonorrhoeae reducing the NO from proinflammatory (>1 microM) to anti-inflammatory (approximately 100 nM) concentrations. The implications of the ability of N. gonorrhoeae to maintain an anti-inflammatory NO concentration is discussed in relation to asymptomatic infection in women.
authors.affiliation: Division of Biology, Alfred University, Alfred, NY 14802, USA.
authors.forename: Jean A
authors.initials: JA
authors.lastname: Cardinale
authors.forename: Virginia L
authors.initials: VL
authors.lastname: Clark
mesh_terms: D000693:Anaerobiosis; D005260:Female; D006069:Gonorrhea / Q000382:microbiology / Q000473:pathology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009344:Neisseria gonorrhoeae / Q000235:genetics / Q000378:metabolism*; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D009572:Nitrite Reductases / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D009609:Nitrous Oxide / Q000378:metabolism; D010088:Oxidoreductases / Q000378:metabolism; D014226:Triazenes / Q000378:metabolism
chemical_list: C105351:1-hydroxy-2-oxo-3,3-bis(2-aminoethyl)-1-triazene; D009573:Nitrites; D014226:Triazenes; D009569:Nitric Oxide; D010088:Oxidoreductases; D009572:Nitrite Reductases; C034747:nitric-oxide reductase; D009609:Nitrous Oxide