Anoxic function for the Escherichia coli flavohaemoglobin (Hmp): reversible binding of nitric oxide and reduction to nitrous oxide.
The flavohaemoglobin Hmp of Escherichia coli is inducible by nitric oxide (NO) and provides protection both aerobically and anaerobically from inhibition of growth by NO and agents that cause nitrosative stress. Here we report rapid kinetic studies of NO binding to Fe(III) Hmp with a second order rate constant of 7.5 x 10(5) M(-1) s(-1) to generate a nitrosyl adduct that was stable anoxically but decayed in the presence of air to reform the Fe(III) protein. NO displaced CO bound to dithionite-reduced Hmp but, remarkably, CO recombined after only 2 s at room temperature indicative of NO reduction and dissociation from the haem. Addition of NO to anoxic NADH-reduced Hmp also generated a nitrosyl species which persisted while NADH was oxidised. These results are consistent with direct demonstration by membrane-inlet mass spectrometry of NO consumption and nitrous oxide production during anoxic incubation of NADH-reduced Hmp. The results demonstrate a new mechanism by which Hmp may eliminate NO under anoxic growth conditions.
Kim
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SO
Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, The University of Sheffield, UK.
Orii
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Y
Lloyd
D
D
Hughes
M N
MN
Poole
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RK
D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000378:metabolism*; D002248:Carbon Monoxide / Q000378:metabolism; D004093:Dihydropteridine Reductase; D004227:Dithionite; D004579:Electron Transport; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins; D006420:Hemeproteins / Q000378:metabolism*; D009247:NADH, NADPH Oxidoreductases; D009569:Nitric Oxide / Q000378:metabolism*; D009609:Nitrous Oxide / Q000378:metabolism*; D010084:Oxidation-Reduction
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006420:Hemeproteins; D004227:Dithionite; D009569:Nitric Oxide; D002248:Carbon Monoxide; D004093:Dihydropteridine Reductase; C076538:hmp protein, E coli; D009247:NADH, NADPH Oxidoreductases; D009609:Nitrous Oxide