Kinetic characterization of the inactivation of ammonia monooxygenase in Nitrosomonas europaea by alkyne, aniline and cyclopropane derivatives.
The kinetic mechanisms of seven inactivators of ammonia oxidation activity in cells of the nitrifying bacterium, Nitrosomonas europaea were investigated. The effects of the inactivators were specific for ammonia monooxygenase (AMO) which oxidizes ammonia to hydroxylamine. The aniline derivatives, 1,3-phenylenediamine and p-anisidine, were potent inactivators of AMO while other derivatives were ineffective as inactivators. Two cyclopropane derivatives, 1, 2-dimethylcyclopropane and cyclopropyl bromide, were inactivators while cyclopropane was not an inactivator. The mechanisms of three alkynes, 1-hexyne, 3-hexyne, and acetylene, were also examined. For all seven compounds, the inactivation of AMO was irreversible, time-dependent, first-order, and dependent on catalytic turnover. Saturation of the rate of inactivation was indicated for p-anisidine (kinact=2.85 min-1; KI=1.0 mM) and cyclopropyl bromide (kinact=4.4 min-1; KI=97 microM), but not for any of the remaining five inactivators, including acetylene. Ammonia slowed the rate of inactivation for acetylene and cyclopropyl bromide, but enhanced the rate of inactivation for the remaining inactivators. All seven compounds appear to be mechanism-based inactivators of AMO.
Keener
W K
WK
Laboratory for Nitrogen Fixation Research, Department of Botany and Plant Pathology, Oregon State University, 2082 Cordley, Corvallis, OR 97331, USA.
Russell
S A
SA
Arp
D J
DJ
D000641:Ammonia / Q000378:metabolism; D000814:Aniline Compounds / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry; D003521:Cyclopropanes / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007700:Kinetics; D015394:Molecular Structure; D009606:Nitrosomonas / Q000201:enzymology*; D010088:Oxidoreductases / Q000737:chemistry*; D010100:Oxygen / Q000378:metabolism; D019163:Reducing Agents / Q000494:pharmacology
D000814:Aniline Compounds; D001426:Bacterial Proteins; D003521:Cyclopropanes; D004791:Enzyme Inhibitors; D019163:Reducing Agents; D000641:Ammonia; D010088:Oxidoreductases; C045582:ammonia monooxygenase; D010100:Oxygen