EC Number | Cloned (Comment) | Organism |
---|---|---|
1.7.1.3 | sequence comparisons, recombinant enzyme expression of wild-type and mutant enzymes in Escherichia coli strains TP1000, RK5206, and RK5204 | Neurospora crassa |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.7.1.3 | G811V | site-directed mutagenesis, an FAD-binding mutant | Neurospora crassa |
1.7.1.3 | H654A/H677A | site-directed mutagenesis, CD spectroscopy shows no negative effects of the introduced mutations on protein secondary structure in comparison to the wild-type protein, but the mutant contains no heme, while the FAD binding ability is not significantly disturbed | Neurospora crassa |
1.7.1.3 | R778E | site-directed mutagenesis, an FAD-binding mutant. The mutant binds essentially the same amount of Moco as does the wild type protein | Neurospora crassa |
1.7.1.3 | Y780A | site-directed mutagenesis, an FAD-binding mutant | Neurospora crassa |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.7.1.3 | additional information | - |
additional information | Michaelis-Menten kinetics, holo-enzyme | Neurospora crassa | |
1.7.1.3 | 0.25 | - |
nitrate | recombinant holo-enzyme, pH 7.5, temperature not specified in the publication | Neurospora crassa |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.7.1.3 | Fe2+ | heme | Neurospora crassa | |
1.7.1.3 | Mo5+ | in the active site-bound molybdenum cofactor | Neurospora crassa |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.7.1.3 | additional information | - |
gel filtration and analytical ultracentrifugation | Neurospora crassa |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.7.1.3 | nitrate + NADPH + H+ | Neurospora crassa | - |
nitrite + NADP+ + H2O | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.7.1.3 | Neurospora crassa | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.7.1.3 | recombinant enzyme from Escherichia coli strains TP1000, RK5206, and RK5204 by a two-step affinity purification | Neurospora crassa |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
1.7.1.3 | 4.32 | - |
purified recombinant holo-enzyme, substrate nitrate, pH 7.5, temperature not specified in the publication | Neurospora crassa |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.7.1.3 | additional information | NADPH-dependent cytochrome c reducing activity by the holo-enzyme is determined with FAD and NADPH spectroscopically at 550 nm and 340 nm. Apo-nitrate reductase has a marginally lower, about 10% reduced cytochrome c reducing activity, which correlates to its 15% reduced heme content | Neurospora crassa | ? | - |
? | |
1.7.1.3 | nitrate + NADPH + H+ | - |
Neurospora crassa | nitrite + NADP+ + H2O | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.7.1.3 | homodimer | holo-enzyme, determination with SDS-PAGE, gel filtration and analytical ultracentrifugation. The enzyme has a largely hydrophobic dimer interface | Neurospora crassa |
1.7.1.3 | More | the apo-enzyme dissociates completely into monomers, the ratio between monomeric and dimeric apo-NR does not change significantly upon a 20fold dilution. Active site formation of eukaryotic nitrate reductase is an autonomous process intrinsically tied to nitrate reductase dimerization, molybdenum cofactor-dependent enzyme maturation, overview. Enzyme domain structure, overview | Neurospora crassa |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.7.1.3 | nitrate reductase [NADPH] | - |
Neurospora crassa |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
1.7.1.3 | 7.2 | - |
assay at | Neurospora crassa |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.7.1.3 | cytochrome c | - |
Neurospora crassa | |
1.7.1.3 | FAD | the enzyme contains a a ferredoxin reductase-type FAD binding domain | Neurospora crassa | |
1.7.1.3 | heme | enzyme heme binding is independent from Moco/MPT binding | Neurospora crassa | |
1.7.1.3 | molybdenum cofactor | i.e Moco/MPT, binding of molybdenum cofactor to apo-nitrate reductase is independent from other prosthetic groups, molybdenum cofactor-dependent enzyme maturation, overview. Reconstitution of Moco-free nitrate reductase with various amounts of purified Moco carrier protein | Neurospora crassa | |
1.7.1.3 | molybdopterin | i.e Moco/MPT, binding of molybdenum cofactor to apo-nitrate reductase is independent from other prosthetic groups, molybdenum cofactor-dependent enzyme maturation, overview. Reconstitution of Moco-free nitrate reductase with various amounts of purified Moco carrier protein | Neurospora crassa | |
1.7.1.3 | NADPH | dependent on | Neurospora crassa |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.7.1.3 | additional information | active site formation of eukaryotic nitrate reductase is an autonomous process intrinsically tied to nitrate reductase dimerization, molybdenum cofactor-dependent enzyme maturation, overview | Neurospora crassa |