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Literature summary extracted from

  • Bennett, K.; Sadler, N.C.; Wright, A.T.; Yeager, C.; Hyman, M.R.
    Activity-based protein profiling of ammonia monooxygenase in Nitrosomonas europaea (2016), Appl. Environ. Microbiol., 82, 2270-2279 .
    View publication on PubMedView publication on EuropePMC

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.99.39 1,7-octadiyne 17OD, complete inhibition, inactivation of NH4+-dependent O2 uptake by Nitrosomonas europaea in a time- and concentration-dependent manner. The effects of 17OD are specific for ammonia-oxidizing activity (17OD has no inhibitory effect on NH2OH-dependent O2 uptake), and de novo protein synthesis is required to reestablish this activity in cells exposed to 17OD. NH4Cl does not protect against inactivation of ammonia-oxidizing activity by 17OD under the conditions tested. 17OD is an irreversible inactivator of AMO Nitrosomonas europaea
1.14.99.39 chloramphenicol rate of NO2- production by 1,7-octadiyne-untreated cells is about 30% lower in the presence of chloramphenicol Nitrosomonas europaea
1.14.99.39 additional information many alkynes are mechanism-based inactivators of AMO, activity-based protein profiling method for the enzyme using 1,7-octadiyneas a probe. Many organic compounds reversibly inhibit ammonia oxidation through their action as alternative substrates for AMO. These compounds include diverse alkanes, alkenes, aromatics, ethers, and halogenated compounds. The simplest organic AMO substrates, such as methane and ethylene, are competitive inhibitors of ammonia oxidation, while other substrates exhibit more complex inhibition patterns. Mechanism-based enzyme inactivation, overview. The free ethynyl group of the inactive enzyme-inactivator adduct is conjugated with either a visualization tag (e.g., Alexa Fluor 647 azide) or an affinity purification tag (e.g., biotin-azide) using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The resulting enzyme-probe-tag conjugant can then either be (i) visualized using IR fluorescence in SDS-PAGE or (ii) enriched by affinity chromatography, tryptically digested, and identified by LC-MS/MS Nitrosomonas europaea
1.14.99.39 rifampicin rate of NO2- production by 1,7-octadiyne-untreated cells is about 15% lower in the presence of rifampin Nitrosomonas europaea

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
1.14.99.39 membrane membrane-associated Nitrosomonas europaea 16020
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.99.39 Cu2+ required Nitrosomonas europaea

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.14.99.39 NH3 + a reduced acceptor + O2 Nitrosomonas europaea
-
 NH2OH + an acceptor + H2O
-
 ?
1.14.99.39 NH3 + a reduced acceptor + O2 Nitrosomonas europaea ATCC 19718
-
 NH2OH + an acceptor + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.14.99.39 Nitrosomonas europaea Q04507 AND Q04508 subunits a and b
-
1.14.99.39 Nitrosomonas europaea ATCC 19718 Q04507 AND Q04508 subunits a and b
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.14.99.39 the free ethynyl group of the inactive enzyme-inactivator adduct is conjugated with either a visualization tag (e.g., Alexa Fluor 647 azide) or an affinity purification tag (e.g., biotin-azide) using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The resulting enzyme-probe-tag conjugant can then either be (i) visualized using IR fluorescence in SDS-PAGE or (ii) enriched by affinity chromatography, tryptically digested, and identified by LC-MS/MS Nitrosomonas europaea

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.99.39 ethylene + a reduced acceptor + O2
-
 Nitrosomonas europaea ? + an acceptor + H2O
-
 ?
1.14.99.39 ethylene + a reduced acceptor + O2
-
 Nitrosomonas europaea ATCC 19718 ? + an acceptor + H2O
-
 ?
1.14.99.39 additional information the free ethynyl group of the inactive enzyme-inactivator adduct is conjugated with either a visualization tag (e.g., Alexa Fluor 647 azide) or an affinity purification tag (e.g., biotin-azide) using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The resulting enzyme-probe-tag conjugant can then either be (i) visualized using IR fluorescence in SDS-PAGE or (ii) enriched by affinity chromatography, tryptically digested, and identified by LC-MS/MS Nitrosomonas europaea ?
-
 ?
1.14.99.39 additional information the free ethynyl group of the inactive enzyme-inactivator adduct is conjugated with either a visualization tag (e.g., Alexa Fluor 647 azide) or an affinity purification tag (e.g., biotin-azide) using a copper-catalyzed azide-alkyne cycloaddition (CuAAC) reaction. The resulting enzyme-probe-tag conjugant can then either be (i) visualized using IR fluorescence in SDS-PAGE or (ii) enriched by affinity chromatography, tryptically digested, and identified by LC-MS/MS Nitrosomonas europaea ATCC 19718 ?
-
 ?
1.14.99.39 NH3 + a reduced acceptor + O2
-
 Nitrosomonas europaea NH2OH + an acceptor + H2O
-
 ?
1.14.99.39 NH3 + a reduced acceptor + O2
-
 Nitrosomonas europaea ATCC 19718 NH2OH + an acceptor + H2O
-
 ?

Subunits

EC Number Subunits Comment Organism
1.14.99.39 ? x * 28000, subunit AmoA, SDS-PAGE Nitrosomonas europaea

Synonyms

EC Number Synonyms Comment Organism
1.14.99.39 AMO
-
 Nitrosomonas europaea
1.14.99.39 amoA
-
 Nitrosomonas europaea

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.14.99.39 30
-
 in vivo assay at Nitrosomonas europaea

General Information

EC Number General Information Comment Organism
1.14.99.39 metabolism Nitrosomonas europaea is an aerobic nitrifying bacterium that oxidizes ammonia (NH3) to nitrite (NO2-) through the sequential activities of ammonia monooxygenase (AMO) and hydroxylamine dehydrogenase (HAO) Nitrosomonas europaea