EC Number | Cloned (Comment) | Organism |
---|---|---|
7.1.1.2 | expression of wild-type and mutant enzymes in Escherichia coli | Escherichia coli |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
7.1.1.2 | E183D | site-directed mutagenesis, the mutant shows a 2fold increase in NADH/ferricyanide oxidoreductase activity and a 5fold increase in NADPHoxidase activity compared to the wild-type enzyme | Escherichia coli |
7.1.1.2 | E183H | site-directed mutagenesis, the mutant shows a 2fold increase in NADH/ferricyanide oxidoreductase activity and a 2fold increase in NADPHoxidase activitycompared to the wild-type enzyme | Escherichia coli |
7.1.1.2 | E183N | site-directed mutagenesis, the mutant shows a 2fold increase in NADH/ferricyanide oxidoreductase activity and a 2fold increase in NADPHoxidase activitycompared to the wild-type enzyme | Escherichia coli |
7.1.1.2 | E183Q | site-directed mutagenesis, the mutant shows a similar NADH/ferricyanide oxidoreductase activity and a 2fold increase NADPHoxidase activity compared to the wild-type enzyme | Escherichia coli |
7.1.1.2 | additional information | construction of several variants with mutations at position 183 exhibiting up to 200fold enhanced catalytic efficiency with NADPH | Escherichia coli |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
7.1.1.2 | Piericidin A | - |
Escherichia coli |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.1.1.2 | 0.0057 | - |
NADH | mutant E183H, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.0058 | - |
NADH | mutant E183D, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.012 | - |
NADH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.013 | - |
NADH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.014 | - |
NADH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.025 | - |
NADPH | mutant E183H, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.045 | - |
NADPH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.39 | - |
NADPH | mutant E183D, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 0.48 | - |
NADPH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 1.87 | - |
NADPH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
7.1.1.2 | membrane | - |
Escherichia coli | 16020 | - |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.1.1.2 | NADH + ubiquinone + 6 H+[side 1] | Escherichia coli | - |
NAD+ + ubiquinol + 7 H+[side 2] | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
7.1.1.2 | Escherichia coli | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
7.1.1.2 | recombinant wild-type and mutant enzymes from Escherichia coli | Escherichia coli |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
7.1.1.2 | additional information | - |
NADH/ferricyanide oxidoreductase activity (measured as ferricyanide reduction) and NAD(P)H oxidase activity (measured as oxygen consumption) of cytoplasmic membranes from Escherichia coli recombinantly expressing the enzyme | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.1.1.2 | NADH + ubiquinone + 6 H+[side 1] | - |
Escherichia coli | NAD+ + ubiquinol + 7 H+[side 2] | - |
? | |
7.1.1.2 | NADPH + ubiquinone + 6 H+[side 1] | NADPH is a poor substrate of the complex | Escherichia coli | NADP+ + ubiquinol + 7 H+[side 2] | - |
? |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
7.1.1.2 | More | the enzyme nucleotide-binding site is made up of a unique Rossmann fold to accommodate the binding of the substrate NADH and of the primary electron acceptor flavin mononucleotide | Escherichia coli |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
7.1.1.2 | complex I | - |
Escherichia coli |
7.1.1.2 | energy-converting NADPH:ubiquinone oxidoreductase | - |
Escherichia coli |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
7.1.1.2 | 30 | - |
assay at | Escherichia coli |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.1.1.2 | 2 | 8 | NADH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 3 | - |
NADPH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 10 | - |
NADPH | mutant E183H, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 11 | - |
NADPH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 11 | - |
NADH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 14 | - |
NADPH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 26 | - |
NADH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 34 | - |
NADPH | mutant E183D, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 37 | - |
NADH | mutants E183D and E18H, pH 6.0, 30°C | Escherichia coli |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
7.1.1.2 | 6 | 7 | mutant E183H, activity with NADH | Escherichia coli |
7.1.1.2 | 6 | - |
mutant E183H, activity with NADPH | Escherichia coli |
7.1.1.2 | 6 | - |
wild-type enzyme, activity with NADH, first peak | Escherichia coli |
7.1.1.2 | 6.5 | - |
wild-type enzyme, activity with NADPH | Escherichia coli |
7.1.1.2 | 7.5 | - |
wild-type enzyme, activity with NADH, second higher peak | Escherichia coli |
EC Number | pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|---|
7.1.1.2 | 5 | 8 | activity range | Escherichia coli |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
7.1.1.2 | FMN | - |
Escherichia coli | |
7.1.1.2 | additional information | NADPH is a poor substrate of the complex | Escherichia coli | |
7.1.1.2 | NADH | binding of NADH includes interactions of the hydroxyl groups of the adenosine ribose with a conserved glutamic acid residue. Structural analysis revealed that due to steric hindrance and electrostatic repulsion, this residue most likely prevents the binding of NADPH, which is a poor substrate of the complex | Escherichia coli |
EC Number | General Information | Comment | Organism |
---|---|---|---|
7.1.1.2 | physiological function | the respiratory complex I couples the electron transfer from NADH to ubiquinone with a translocation of protons across the membrane | Escherichia coli |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
7.1.1.2 | 1.8 | - |
NADPH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 20 | - |
NADPH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 90 | - |
NADPH | mutant E183D, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 320 | - |
NADPH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 400 | - |
NADPH | mutant E183H, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 800 | - |
NADH | mutant E183N, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 2000 | - |
NADH | wild-type enzyme, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 2300 | - |
NADH | mutant E183Q, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 6400 | - |
NADH | mutant E183D, pH 6.0, 30°C | Escherichia coli | |
7.1.1.2 | 6500 | - |
NADH | mutant E183H, pH 6.0, 30°C | Escherichia coli |