Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Anabaena sp. |
Protein Variants | Comment | Organism |
---|---|---|
Y303F | site-directed mutagenesis, about 30% of the wild-type enzyme activity with ferredoxin, about 25% of the wild-type enzyme activity with flavodoxin | Anabaena sp. |
Y303S | site-directed mutagenesis, inactive mutant | Anabaena sp. |
Y303W | site-directed mutagenesis, nearly inactive mutant | Anabaena sp. |
Y308F | site-directed mutagenesis, about 20% of the wild-type enzyme activity with ferredoxin, about 11% of the wild-type enzyme activity with flavodoxin | Pisum sativum |
Y308S | site-directed mutagenesis, about 5% of the wild-type enzyme activity with ferredoxin, no activity with flavodoxin | Pisum sativum |
Y308W | site-directed mutagenesis, nearly inactive mutant with ferredoxin, about 25% of the wild-type enzyme activity with flavodoxin | Pisum sativum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | stopped-flow and laser flash kinetic measurements, steady-state kinetics, dissociation constants and reduction potentials of wild-type and mutant enzymes | Pisum sativum | |
additional information | - |
additional information | stopped-flow and laser flash kinetic measurements, steady-state kinetics, dissociation constants and reduction potentials of wild-type and mutant enzymes | Anabaena sp. | |
0.0058 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308F | Pisum sativum | |
0.0065 | - |
reduced ferredoxin | pH 8.0, 13°C, wild-type enzyme | Pisum sativum | |
0.009 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308S | Pisum sativum | |
0.011 | - |
reduced ferredoxin | pH 8.0, 13°C, wild-type enzyme | Anabaena sp. | |
0.017 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308W | Pisum sativum | |
0.051 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y303F | Anabaena sp. |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Anabaena sp. | P21890 | - |
- |
Pisum sativum | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) | Anabaena sp. |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH | catalytic mechanism, initiated by reduction of FAD cofactor by obligatory one-electron carriers ferredoxin or flavodoxin in presence of NADP+, the enzymes' C-terminal tyrosine residue is involved modulating the enzyme affinity for NADP+/NADPH | Pisum sativum | |
2 reduced ferredoxin + NADP+ + H+ = 2 oxidized ferredoxin + NADPH | catalytic mechanism, initiated by reduction of FAD cofactor by obligatory one-electron carriers ferredoxin or flavodoxin in presence of NADP+, the enzymes' C-terminal tyrosine residue is involved modulating the enzyme affinity for NADP+/NADPH | Anabaena sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
reduced ferredoxin + NADP+ | - |
Pisum sativum | oxidized ferredoxin + NADPH | - |
r | |
reduced ferredoxin + NADP+ | - |
Anabaena sp. | oxidized ferredoxin + NADPH | - |
r | |
reduced ferredoxin + NADP+ | - |
Pisum sativum | oxidized ferredoxin + NADPH + H+ | - |
r | |
reduced ferredoxin + NADP+ | - |
Anabaena sp. | oxidized ferredoxin + NADPH + H+ | - |
r | |
reduced flavodoxin + NADP+ | - |
Pisum sativum | oxidized flavodoxin + NADPH + H+ | - |
r | |
reduced flavodoxin + NADP+ | - |
Anabaena sp. | oxidized flavodoxin + NADPH + H+ | - |
r |
Synonyms | Comment | Organism |
---|---|---|
ferredoxin-nicotinamide adenine dinucleotide phosphate reductase | - |
Pisum sativum |
ferredoxin-nicotinamide adenine dinucleotide phosphate reductase | - |
Anabaena sp. |
FNR | - |
Pisum sativum |
FNR | - |
Anabaena sp. |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
13 | - |
assay at | Pisum sativum |
13 | - |
assay at | Anabaena sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y303W | Anabaena sp. | |
2.5 | - |
reduced flavodoxin | pH 8.0, 13°C, mutant Y303W | Anabaena sp. | |
2.5 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308W | Pisum sativum | |
4 | - |
reduced flavodoxin | pH 8.0, 13°C, mutant Y308F | Pisum sativum | |
7 | - |
reduced flavodoxin | pH 8.0, 13°C, mutant Y303F | Anabaena sp. | |
7.7 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308S | Pisum sativum | |
8.3 | - |
reduced flavodoxin | pH 8.0, 13°C, mutant Y308W | Pisum sativum | |
23.3 | - |
reduced flavodoxin | pH 8.0, 13°C, wild-type enzyme | Anabaena sp. | |
23.9 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y308F | Pisum sativum | |
30.6 | - |
reduced flavodoxin | pH 8.0, 13°C, wild-type enzyme | Pisum sativum | |
32 | - |
reduced ferredoxin | pH 8.0, 13°C, mutant Y303F | Anabaena sp. | |
139 | - |
reduced ferredoxin | pH 8.0, 13°C, wild-type enzyme | Pisum sativum | |
200 | - |
reduced ferredoxin | pH 8.0, 13°C, wild-type enzyme | Anabaena sp. |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Pisum sativum |
8 | - |
assay at | Anabaena sp. |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | binding and hydride/electron transfer mechanism | Pisum sativum | |
FAD | binding and hydride/electron transfer mechanism | Anabaena sp. | |
NADP+ | binding and hydride transfer mechanism | Pisum sativum | |
NADP+ | binding and hydride transfer mechanism | Anabaena sp. | |
NADPH | binding and hydride transfer mechanism | Pisum sativum | |
NADPH | binding and hydride transfer mechanism | Anabaena sp. |