Literature summary for 1.7.2.1 extracted from

Tocheva, E.I.; Eltis, L.D.; Murphy, M.E.
Conserved active site residues limit inhibition of a copper-containing nitrite reductase by small molecules (2008), Biochemistry, 47, 4452-4460.

Cloned(Commentary)

Cloned (Comment)	Organism
expression of the wild-type enzyme in Escherichia coli strain BL21 (DE3)	Alcaligenes faecalis

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme, free or in complex with small molecule inhibitors, hanging drop vapor diffusion method, room temperature, 25 mg/ml protein in 20 mM Tris-HCl, pH 7.0, is mixed with an equal volume of reservoir containing 6-10% PEG 4000, 100 mM sodium acetate, pH 4.0, addition of 20 mM of ligands 20 mM of azide, formate, or nitrate, X-ray diffraction structure determination and analysis at 1.5-1.8 A resolution	Alcaligenes faecalis

Crystallization (Comment)

Organism

purified recombinant enzyme, free or in complex with small molecule inhibitors, hanging drop vapor diffusion method, room temperature, 25 mg/ml protein in 20 mM Tris-HCl, pH 7.0, is mixed with an equal volume of reservoir containing 6-10% PEG 4000, 100 mM sodium acetate, pH 4.0, addition of 20 mM of ligands 20 mM of azide, formate, or nitrate, X-ray diffraction structure determination and analysis at 1.5-1.8 A resolution

Alcaligenes faecalis

Inhibitors

Inhibitors	Comment	Organism
acetate	weak, mixed-type inhibition, inhibition mode, overview	Alcaligenes faecalis
azide	binding mode, overview	Alcaligenes faecalis
formate	weak mixed-type inhibition, inhibition mode, overview	Alcaligenes faecalis
nitrate	weak inhibition, inhibition mode, overview	Alcaligenes faecalis
nitrous oxide	binding mode, overview	Alcaligenes faecalis

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
additional information	-	additional information	steady-state kinetics	Alcaligenes faecalis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Cu2+	copper-containing dissimilatory nitrite reductase, catalytic type 2 copper, binding site structure, analysis of binding structure and interaction with inhibitors, overview	Alcaligenes faecalis

Organism

Organism	UniProt	Comment	Textmining
Alcaligenes faecalis	P38501	-	-
Alcaligenes faecalis S-6	P38501	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant wild-type enzyme from Escherichia coli strain BL21 (DE3) by nickel affinity chromatography, the tag is removed by thrombin, followed by anion exchange chromatography	Alcaligenes faecalis

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
additional information	the active site residue is Ile257. The small molecules formate, acetate and nitrate mimic the substrate by having at least two oxygen atoms for bidentate coordination to the type 2 copper atom and interacting wit the oxidized catalytic metal ion, overview. Nitrite and the substrate mimic bind in the same asymmetric, bidentate manner	Alcaligenes faecalis	?	-	?
additional information	the active site residue is Ile257. The small molecules formate, acetate and nitrate mimic the substrate by having at least two oxygen atoms for bidentate coordination to the type 2 copper atom and interacting wit the oxidized catalytic metal ion, overview. Nitrite and the substrate mimic bind in the same asymmetric, bidentate manner	Alcaligenes faecalis S-6	?	-	?
nitrite + H2O + reduced pseudoazurin	reduction of pseudoazurin by ascorbate	Alcaligenes faecalis	nitric oxide + H+ + pseudoazurin	-	?
nitrite + H2O + reduced pseudoazurin	reduction of pseudoazurin by ascorbate	Alcaligenes faecalis S-6	nitric oxide + H+ + pseudoazurin	-	?

Synonyms

Synonyms	Comment	Organism
copper-containing dissimilatory nitrite reductase	-	Alcaligenes faecalis
copper-containing nitrite reductase	-	Alcaligenes faecalis
NiR	-	Alcaligenes faecalis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Alcaligenes faecalis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
4	6.5	assay at, pH dependence, overview	Alcaligenes faecalis

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
2	-	azide	recombinant enzyme, pH 6.5, 25°C	Alcaligenes faecalis