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Literature summary extracted from

  • Hough, M.A.; Eady, R.R.; Hasnain, S.S.
    Identification of the proton channel to the active site type 2 Cu center of nitrite reductase: structural and enzymatic properties of the His254Phe and Asn90Ser mutants (2008), Biochemistry, 47, 13547-13553.
    View publication on PubMed

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.7.2.1 mutant H254F loaded with either Cu2+ or Zn2+, to 1.5 A and 1.85 A, respectively. Both structures are essentially identical. Structure of mutant N90S, to 1.6 A resolution. In both the native and mutant N90S structures, a surface Zn ion is present in each monomer, bridging the two monomers through the coordinating residues His165 and Asp167 of one monomer and Glu195 of the adjacent monomer. This Zn site is similar to that described previously in several NiR structures Achromobacter xylosoxidans

Protein Variants

EC Number Protein Variants Comment Organism
1.7.2.1 H254F full catalytic activity despite disruption of the primary proton channel. No change in apparent Km value for nitrite Achromobacter xylosoxidans
1.7.2.1 N90S disruption of H-bonding in the high-pH proton channel results in an 70% decrease in specific activity. No change in apparent Km value for nitrite Achromobacter xylosoxidans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.7.2.1 0.034
-
 nitrite wild-type, pH 7.1 Achromobacter xylosoxidans
1.7.2.1 0.05
-
 nitrite mutant H254F, pH 7.1 Achromobacter xylosoxidans
1.7.2.1 0.05
-
 nitrite mutant N90S, pH 7.1 Achromobacter xylosoxidans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.7.2.1 Zn2+ present on surface of each monomer, crystallization data Achromobacter xylosoxidans

Organism

EC Number Organism UniProt Comment Textmining
1.7.2.1 Achromobacter xylosoxidans O68601
-
-

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.7.2.1 82
-
 mutant N90S, pH 7.1 Achromobacter xylosoxidans
1.7.2.1 123
-
 mutant H254F, pH 7.1 Achromobacter xylosoxidans
1.7.2.1 240
-
 wild-type, pH 7.1 Achromobacter xylosoxidans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.7.2.1 nitrite + reduced azurin I
-
 Achromobacter xylosoxidans NO + azurin I
-
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