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Literature summary extracted from

  • Cutruzzola, F.; Brown, K.; Wilson, E.K.; Bellelli, A.; Arese, M.; Tegoni, M.
    The nitrite reductase from Pseudomonas aeruginosa: essential role of two active-site histidines in the catalytic and structural properties (2001), Proc. Natl. Acad. Sci. USA, 98, 2232-2237.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.7.2.1 expression of wild-type, H327A and H369A mutant enzyme in Pseudomonas putida Pseudomonas aeruginosa

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.7.2.1 crystals of the H327A mutant are obtained by vapor diffusion technique by mixing in a 1:1 ratio the protein and a reservoir solution containing 4.0% polyethylene glycol 5000 monomethyl ether, 0.1 M sodium acetate, pH 5.5. The space group is 4(3)22 with cell dimensions 70.5 x 70.5 x 281 A. Crystals of the H369A mutant are obtained by mixing in a 1.1 ratio the protein and a reservoir solution containing 11.5% polyethylene glycol 6000, 0.2 M imidazole/malate, pH 6.5. The space group is P4(1)2(1)2 with cell dimensions 94.7 x 94.7 x 159.9 A Pseudomonas aeruginosa
1.7.2.1 H327A mutant enzymes: vapour diffusion technique, mixing of the enzyme and a reservoir solution containing 4% polyethylene glycol 5000 monomethyl ether, 100 mM sodium acetate pH 5.5 in a 1/1 ratio, H369A mutant enzyme: 11.5% polyethylene glycol 6000, 200 mM imidazole/malate pH 6.5, x-ray structure of both mutants Pseudomonas aeruginosa

Protein Variants

EC Number Protein Variants Comment Organism
1.7.2.1 H327A reduction of nitrite is severely compromised Pseudomonas aeruginosa
1.7.2.1 H327A mutant protein has no nitrite reductase activity but maintains the ability to reduce O2 to water. Nitrite reductase activity is impaired because of the accumulation of a catalytically inactive form Pseudomonas aeruginosa
1.7.2.1 H369A reduction of nitrite is severely compromised Pseudomonas aeruginosa
1.7.2.1 H369A mutant protein has no nitrite reductase activity but maintains the ability to reduce O2 to water. Nitrite reductase activity is impaired because of the accumulation of a catalytically inactive form Pseudomonas aeruginosa

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.7.2.1 0.0018
-
ferricytochrome c551 recombinant H327A mutant enzyme, O2 reduction Pseudomonas aeruginosa
1.7.2.1 0.0018
-
Ferrocytochrome c-551 pH 6.2, 25°C, mutant enzyme H327A Pseudomonas aeruginosa
1.7.2.1 0.002
-
ferricytochrome c551 recombinant wild-type enzyme, O2 reduction Pseudomonas aeruginosa
1.7.2.1 0.002
-
Ferrocytochrome c-551 pH 6.2, 25°C, wild-type enzyme Pseudomonas aeruginosa
1.7.2.1 0.006
-
NO2- recombinant wild-type enzyme Pseudomonas aeruginosa
1.7.2.1 0.006
-
nitrite pH 6.2, 25°C, wild-type enzyme Pseudomonas aeruginosa
1.7.2.1 0.0075
-
ferricytochrome c551 recombinant H369A mutant enzyme, O2 reduction Pseudomonas aeruginosa
1.7.2.1 0.0075
-
Ferrocytochrome c-551 pH 6.2, 25°C, mutant enzyme H369A Pseudomonas aeruginosa

Organism

EC Number Organism UniProt Comment Textmining
1.7.2.1 Pseudomonas aeruginosa P24474
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.7.2.1 recombinant wild-type, H327A and H369A mutant enzyme Pseudomonas aeruginosa

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.7.2.1 ferrocytochrome c-551 + O2
-
Pseudomonas aeruginosa ferricytochrome c-551 + H2O
-
?
1.7.2.1 nitric oxide + H2O + ferricytochrome c551
-
Pseudomonas aeruginosa nitrite + ferrocytochrome c551 + H+
-
r
1.7.2.1 nitrite + electron donor
-
Pseudomonas aeruginosa NO + oxidized electron donor + H2O
-
?

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.7.2.1 0.025
-
Ferrocytochrome c-551 pH 6.2, 25°C, wild-type enzyme Pseudomonas aeruginosa
1.7.2.1 0.032
-
Ferrocytochrome c-551 pH 6.2, 25°C, mutant enzyme H369A Pseudomonas aeruginosa
1.7.2.1 0.043
-
Ferrocytochrome c-551 pH 6.2, 25°C, mutant enzyme H327A Pseudomonas aeruginosa
1.7.2.1 0.08
-
NO2- recombinant H327A and H369a mutant enzymes Pseudomonas aeruginosa
1.7.2.1 0.08
-
nitrite pH 6.2, 25°C, mutant enzyme H327A Pseudomonas aeruginosa
1.7.2.1 0.08
-
nitrite pH 6.2, 25°C, mutant enzyme H369A Pseudomonas aeruginosa
1.7.2.1 8
-
NO2- recombinant wild-type enzyme Pseudomonas aeruginosa
1.7.2.1 8
-
nitrite pH 6.2, 25°C, wild-type enzyme Pseudomonas aeruginosa

Cofactor

EC Number Cofactor Comment Organism Structure
1.7.2.1 Heme d1 reduction of the substrate occurs at the d1-heme site Pseudomonas aeruginosa