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Literature summary for 1.7.1.3 extracted from

  • Pan, S.S.; Nason, A.
    Purification and characterization of homogeneous assimilatory reduced nicotinamide adenine dinucleotide phosphate-nitrate reductase from Neurospora crassa (1978), Biochim. Biophys. Acta, 523, 297-313.
    View publication on PubMed

Activating Compound

Activating Compound Comment Organism Structure
o-phenanthroline 40% increase of activity at 2.5 mM Neurospora crassa

Inhibitors

Inhibitors Comment Organism Structure
8-hydroxyquinoline 94% inhibition at 10 mM Neurospora crassa
azide 96% inhibition at 5 mM Neurospora crassa
cyanide
-
Neurospora crassa
EDTA
-
Neurospora crassa
additional information protection of inactivation by FAD and restorage by dithiothreitol Neurospora crassa
p-hydroxymercuribenzoate reversed by sulfhydryl reagents Neurospora crassa
Phenylglyoxal in 0.1 M phosphate, pH 7.3, 4 mM, inactivation after 15 min to 40% and to 20% after 60 min Neurospora crassa

Metals/Ions

Metals/Ions Comment Organism Structure
Molybdenum 1 mol per mol protein Neurospora crassa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
115000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested Neurospora crassa
130000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested Neurospora crassa
150000
-
1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested Neurospora crassa
230000
-
sucrose density gradient, gel filtration Neurospora crassa

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
nitrate + NADPH Neurospora crassa first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents nitrite + NADP+ + H2O
-
?
nitrate + NADPH Neurospora crassa STA4 first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents nitrite + NADP+ + H2O
-
?

Organic Solvent Stability

Organic Solvent Comment Organism
Ethanol mild denaturation Neurospora crassa

Organism

Organism UniProt Comment Textmining
Neurospora crassa
-
STA4
-
Neurospora crassa STA4
-
STA4
-

Purification (Commentary)

Purification (Comment) Organism
streptomycin sulfate and ammonium sulfate precipitation, ion-exchange, gel filtration, isoelectric focusing, FAD-affinity Neurospora crassa

Source Tissue

Source Tissue Comment Organism Textmining
mycelium
-
Neurospora crassa
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [Āµmol/min/mg] Specific Activity Maximum [Āµmol/min/mg] Comment Organism
additional information
-
-
Neurospora crassa

Storage Stability

Storage Stability Organism
-10Ā°C, 50 mM sodium phosphate, pH 6.9, 30% glycerol, 0.5 mM EDTA, 0.5 mM dithiothreitol, 0.5 mM phenylmethylsulfonylfluoride, 0.1 mM FAD, stable for at least 6 months Neurospora crassa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
nitrate + NADPH 4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase Neurospora crassa nitrite + NADP+ + H2O
-
?
nitrate + NADPH first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents Neurospora crassa nitrite + NADP+ + H2O
-
?
nitrate + NADPH 4 activities: NADPH-nitrate reductase, FADH-nitrate reductase, reduced methyl viologen-nitrate reductase and NADPH-cytochrome c reductase Neurospora crassa STA4 nitrite + NADP+ + H2O
-
?
nitrate + NADPH first step in the reduction of nitrate to ammonia, biosynthesis of amino acids and other nitrogen-containing cell constituents Neurospora crassa STA4 nitrite + NADP+ + H2O
-
?

Subunits

Subunits Comment Organism
dimer 1 * 130000 + 1 * 115000, SDS-PAGE, homodimer of 2 * 150000 suggested Neurospora crassa

Cofactor

Cofactor Comment Organism Structure
cytochrome b557 heme-iron, 2 mol per mol enzyme Neurospora crassa
FAD 1 mol per subunit is assumed Neurospora crassa