EC Number | Cloned (Comment) | Organism |
---|---|---|
1.6.2.2 | expressed in Escherichia coli BL21(DE3) cells | Sus scrofa |
1.6.2.2 | soluble domain, His-tag | Sus scrofa |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
1.6.2.2 | P247A | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P247A | the mutant shows increased Km and decreased kcat values for NADH and cytochrome b5, as well as increased Km and kcat values for ferricyanide compared to the wild type enzyme | Sus scrofa |
1.6.2.2 | P247L | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P247L | the mutation significantly decreases kcat with slight increase (about 2fold) in Km for the physiological electron donor NADH. However, Km and kcat values for the electron acceptors (both cytochrome b5 and ferricyanide) are decreased significantly | Sus scrofa |
1.6.2.2 | P248A | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P248A | the mutant shows increased Km and decreased kcat values for NADH and cytochrome b5, as well as increased Km and kcat values for ferricyanide compared to the wild type enzyme | Sus scrofa |
1.6.2.2 | P248L | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P248L | the mutant shows increased Km and decreased kcat values for NADH and ferricyanide compared to the wild type enzyme | Sus scrofa |
1.6.2.2 | P249A | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P249A | the mutation affects the Km (NADH) values to increase slightly by a factor of 3 compared to the wild type enzyme | Sus scrofa |
1.6.2.2 | P249L | proposed NADH-binding site, soluble domain is analyzed | Sus scrofa |
1.6.2.2 | P249L | the mutant shows increased Km and kcat values for NADH, ferricyanide and cytochrome b5 compared to the wild type enzyme | Sus scrofa |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.6.2.2 | 0.00104 | - |
ferricyanide | mutant P247L | Sus scrofa | |
1.6.2.2 | 0.00104 | - |
ferricyanide | mutant enzyme P247L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.00693 | - |
ferricytochrome b5 | mutant P247L | Sus scrofa | |
1.6.2.2 | 0.00723 | - |
ferricyanide | - |
Sus scrofa | |
1.6.2.2 | 0.00723 | - |
ferricyanide | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01096 | - |
ferricyanide | mutant P247A | Sus scrofa | |
1.6.2.2 | 0.01096 | - |
ferricyanide | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01097 | - |
NADH | ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.01097 | - |
NADH | wild type enzyme, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.0135 | - |
NADH | mutant P248L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.0135 | - |
NADH | mutant enzyme P248L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01353 | - |
ferricytochrome b5 | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.0155 | - |
NADH | mutant P249L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.0155 | - |
NADH | mutant enzyme P249L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01563 | - |
ferricyanide | mutant P248A | Sus scrofa | |
1.6.2.2 | 0.01563 | - |
ferricyanide | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01568 | - |
ferricyanide | mutant P248L | Sus scrofa | |
1.6.2.2 | 0.01568 | - |
ferricyanide | mutant enzyme P248L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01713 | - |
NADH | mutant P247A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.01713 | - |
NADH | mutant enzyme P247A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.01735 | - |
ferricytochrome b5 | - |
Sus scrofa | |
1.6.2.2 | 0.0207 | - |
ferricytochrome b5 | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.02407 | - |
ferricytochrome b5 | mutant P247A | Sus scrofa | |
1.6.2.2 | 0.02407 | - |
NADH | mutant P248A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.02407 | - |
NADH | mutant enzyme P248A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.02467 | - |
NADH | mutant P247L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.02467 | - |
NADH | mutant enzyme P247L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.02646 | - |
ferricyanide | mutant P249A | Sus scrofa | |
1.6.2.2 | 0.02646 | - |
ferricyanide | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.02928 | - |
NADH | mutant P249A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 0.02928 | - |
NADH | mutant enzyme P249A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.03309 | - |
ferricyanide | mutant P249L | Sus scrofa | |
1.6.2.2 | 0.03309 | - |
ferricyanide | mutant enzyme P249L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.03434 | - |
ferricytochrome b5 | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.04148 | - |
ferricytochrome b5 | mutant P248A | Sus scrofa | |
1.6.2.2 | 0.04481 | - |
ferricytochrome b5 | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 0.05238 | - |
ferricytochrome b5 | mutant P249A | Sus scrofa | |
1.6.2.2 | 0.055 | - |
ferricytochrome b5 | mutant P249L | Sus scrofa | |
1.6.2.2 | 0.08199 | - |
ferricytochrome b5 | mutant P248L | Sus scrofa |
EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|---|
1.6.2.2 | 30810 | - |
calculated from amino acid sequence | Sus scrofa |
1.6.2.2 | 31000 | - |
SDS-PAGE | Sus scrofa |
1.6.2.2 | 33000 | - |
x * 33000 (soluble domain), SDS-PAGE | Sus scrofa |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.6.2.2 | 2 ferricytochrome b5 + NADH | Sus scrofa | - |
2 ferrocytochrome b5 + NAD+ + H+ | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.6.2.2 | Sus scrofa | - |
- |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
1.6.2.2 | ammonium sulfate precipitation, DEAE-Sepharose CL-6B column chromatography, 5'-ADP-agarose (or 5'-AMP-agarose) gel column chromatography, and Ni-NTA-agarose column chromatography | Sus scrofa |
1.6.2.2 | soluble domain | Sus scrofa |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.6.2.2 | 2 ferricyanide + NADH | - |
Sus scrofa | 2 ferrocyanide + NAD+ + H+ | - |
? | |
1.6.2.2 | 2 ferricytochrome b5 + NADH | - |
Sus scrofa | 2 ferrocytochrome b5 + NAD+ + H+ | - |
? | |
1.6.2.2 | NADH + ferricytochrome b5 | - |
Sus scrofa | NAD+ + H+ + ferrocytochrome b5 | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
1.6.2.2 | ? | x * 33000 (soluble domain), SDS-PAGE | Sus scrofa |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.6.2.2 | B5R | - |
Sus scrofa |
1.6.2.2 | NADH-cytochrome b5 reductase | - |
Sus scrofa |
1.6.2.2 | NADH-ferricyanide reductase | - |
Sus scrofa |
1.6.2.2 | NADH-ferricytochrome reductase | - |
Sus scrofa |
1.6.2.2 | NADH:cytochrome b5 reductase | - |
Sus scrofa |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.6.2.2 | 21.7 | - |
NADH | mutant P247L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 21.7 | - |
NADH | mutant enzyme P247L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 27.2 | - |
ferricytochrome b5 | mutant P247L | Sus scrofa | |
1.6.2.2 | 30.8 | - |
ferricyanide | mutant P247L | Sus scrofa | |
1.6.2.2 | 30.8 | - |
ferricyanide | mutant enzyme P247L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 147 | - |
ferricytochrome b5 | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 167 | - |
ferricytochrome b5 | - |
Sus scrofa | |
1.6.2.2 | 167 | - |
ferricytochrome b5 | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 200 | - |
ferricytochrome b5 | mutant P247A | Sus scrofa | |
1.6.2.2 | 270 | - |
ferricytochrome b5 | mutant P249L | Sus scrofa | |
1.6.2.2 | 275 | - |
ferricytochrome b5 | mutant P248L | Sus scrofa | |
1.6.2.2 | 288 | - |
ferricytochrome b5 | mutant P248A | Sus scrofa | |
1.6.2.2 | 301 | - |
ferricytochrome b5 | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 337 | - |
ferricyanide | - |
Sus scrofa | |
1.6.2.2 | 337 | - |
ferricyanide | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 354 | - |
NADH | mutant P247A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 354 | - |
NADH | mutant enzyme P247A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 357 | - |
ferricyanide | mutant P247A | Sus scrofa | |
1.6.2.2 | 357 | - |
ferricyanide | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 399 | - |
ferricyanide | mutant P248A | Sus scrofa | |
1.6.2.2 | 399 | - |
ferricyanide | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 441 | - |
ferricytochrome b5 | mutant P249A | Sus scrofa | |
1.6.2.2 | 504 | - |
ferricyanide | mutant P248L | Sus scrofa | |
1.6.2.2 | 504 | - |
ferricyanide | mutant enzyme P248L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 540 | - |
NADH | mutant P248L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 540 | - |
NADH | mutant enzyme P248L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 540 | - |
ferricytochrome b5 | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 559 | - |
NADH | mutant P248A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 559 | - |
NADH | mutant enzyme P248A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 681 | - |
NADH | mutant P249A, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 681 | - |
NADH | mutant enzyme P249A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 742 | - |
NADH | ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 742 | - |
NADH | wild type enzyme, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 838 | - |
ferricyanide | mutant P249A | Sus scrofa | |
1.6.2.2 | 838 | - |
ferricyanide | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 1059 | - |
NADH | mutant P249L, ferricyanide as electron acceptor | Sus scrofa | |
1.6.2.2 | 1059 | - |
NADH | mutant enzyme P249L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 1241 | - |
ferricyanide | mutant P249L | Sus scrofa | |
1.6.2.2 | 1241 | - |
ferricyanide | mutant enzyme P249L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
1.6.2.2 | FAD | - |
Sus scrofa | |
1.6.2.2 | NADH | - |
Sus scrofa |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.6.2.2 | 880 | - |
NADH | mutant enzyme P247L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 20670 | - |
NADH | mutant enzyme P247A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 23220 | - |
NADH | mutant enzyme P248A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 23260 | - |
NADH | mutant enzyme P249A, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 25530 | - |
ferricyanide | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 29620 | - |
ferricyanide | mutant enzyme P247L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 31670 | - |
ferricyanide | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 32140 | - |
ferricyanide | mutant enzyme P248L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 32570 | - |
ferricyanide | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 37500 | - |
ferricyanide | mutant enzyme P249L, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 40000 | - |
NADH | mutant enzyme P248L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 46610 | - |
ferricyanide | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 67640 | - |
NADH | wild type enzyme, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 68320 | - |
NADH | mutant enzyme P249L, using ferricyanide as cosubstrate, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 80700 | - |
ferricytochrome b5 | mutant enzyme P247A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 87700 | - |
ferricytochrome b5 | mutant enzyme P248A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 108600 | - |
ferricytochrome b5 | wild type enzyme, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa | |
1.6.2.2 | 120500 | - |
ferricytochrome b5 | mutant enzyme P249A, in 100 mM potassium phosphate buffer (pH 7.0) at 25°C | Sus scrofa |