Cloned (Comment) | Organism |
---|---|
expression of wild-type, K110R, K110H, K110A, K110E and K110Q mutant enzymes in Escherichia coli | Rattus norvegicus |
Crystallization (Comment) | Organism |
---|---|
sitting drop method, reservoir: 8% poly ethylene glycol 6000, 5% 2-methyl-2,4-pentanediol in 100 mM 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid, pH 7.5, X-ray structure, resolution: enzyme 2.0 A, enzyme-NAD+ complex, 2.3 A | Rattus norvegicus |
Protein Variants | Comment | Organism |
---|---|---|
K110A | strongly reduced kcat for ferricyanide and cytochrome b5 | Rattus norvegicus |
K110E | strongly reduced kcat for ferricyanide and cytochrome b5 | Rattus norvegicus |
K110H | strongly reduced kcat for ferricyanide and cytochrome b5 | Rattus norvegicus |
K110Q | very low kcat for ferricyanide and cytochrome b5 | Rattus norvegicus |
K110R | reduced kcat for ferricyanid and cytochrome b5 | Rattus norvegicus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
cytochrome b5 | K110Q mutant enzyme | Rattus norvegicus | |
0.004 | - |
cytochrome b5 | K110E mutant enzyme | Rattus norvegicus | |
0.005 | - |
ferricyanide | K110R mutant enzyme | Rattus norvegicus | |
0.006 | - |
ferricyanide | recombinant wild-type enzyme | Rattus norvegicus | |
0.009 | - |
cytochrome b5 | K110R mutant enzyme | Rattus norvegicus | |
0.009 | - |
cytochrome b5 | K110A mutant enzyme | Rattus norvegicus | |
0.01 | - |
cytochrome b5 | recombinant wild-type enzyme | Rattus norvegicus | |
0.012 | - |
cytochrome b5 | K110H mutant enzyme | Rattus norvegicus | |
0.05 | - |
ferricyanide | K110H mutant enzyme | Rattus norvegicus | |
0.37 | - |
ferricyanide | K110E mutant enzyme | Rattus norvegicus | |
0.38 | - |
ferricyanide | K110A mutant enzyme | Rattus norvegicus | |
0.76 | - |
ferricyanide | K110Q mutant enzyme | Rattus norvegicus |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rattus norvegicus | P20070 | - |
- |
Purification (Comment) | Organism |
---|---|
wild-type, K110R, K110H, K110A, K110E and K110Q mutant enzymes | Rattus norvegicus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
NADH + ferricytochrome b5 | - |
Rattus norvegicus | NAD+ + H+ + ferrocytochrome b5 | - |
r |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
ferricytochrome b5 | K110E mutant enzyme | Rattus norvegicus | |
4 | - |
ferricytochrome b5 | K110Q mutant enzyme | Rattus norvegicus | |
40 | - |
ferricyanide | K110Q mutant enzyme | Rattus norvegicus | |
90 | - |
ferricytochrome b5 | K110A mutant enzyme | Rattus norvegicus | |
110 | - |
ferricytochrome b5 | K110H mutant enzyme | Rattus norvegicus | |
120 | - |
ferricyanide | K110E mutant enzyme | Rattus norvegicus | |
200 | - |
ferricytochrome b5 | K110R mutant enzyme | Rattus norvegicus | |
270 | - |
ferricytochrome b5 | recombinant wild-type enzyme | Rattus norvegicus | |
270 | - |
ferricyanide | K110A mutant enzyme | Rattus norvegicus | |
340 | - |
ferricyanide | K110H mutant enzyme | Rattus norvegicus | |
470 | - |
ferricyanide | K110R mutant enzyme | Rattus norvegicus | |
800 | - |
ferricyanide | recombinant wild-type enzyme | Rattus norvegicus |