Cloned (Comment) | Organism |
---|---|
gene fccA, expression of wild-type and mutant flavocytochrome enzymes in the enzyme-deficient strain DELTAfccA | Shewanella frigidimarina |
Crystallization (Comment) | Organism |
---|---|
mutant enzyme H505A and H505Y, hanging drop vapour diffusion method, protein solution: 6 mg/ml, 10 mM Tris-HCl, pH 8.5, well solution: 100 mM Tris-HCl, pH 7.4-8.2, 25°C, 80 mM NaCl, 16-19% PEG 8000, 10 mM fumarate, equal volume of 0.002 ml of protein solution and well solution, 10 days, cryoprotectant solution: 100 mM sodium acetate, pH 6.5, 20% PEG 8000, 10 mM fumarate, 80 mM NaCl, and 23% glycerol, X-ray diffraction structure determination and analysis at 1.8 A and 2.0 A resolution, respectively, molecular replacement | Shewanella frigidimarina |
Protein Variants | Comment | Organism |
---|---|---|
H505A | site-directed mutagenesis, altered binding structure of the sodium ion, crystal structure analysis, at pH 8.5 the activity of the mutant enzyme is similar to the wild-type enzyme, at pH 6.0 the ctivity is 20fold reduced | Shewanella frigidimarina |
H505Y | site-directed mutagenesis, altered binding structure of the sodium ion, crystal structure analysis, at pH 8.5 the activity of the mutant enzyme is similar to the wild-type enzyme | Shewanella frigidimarina |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.007 | - |
fumarate | wild-type enzyme, pH 9.0, 25°C | Shewanella frigidimarina | |
0.009 | - |
fumarate | mutant H505A, pH 9.0, 25°C | Shewanella frigidimarina | |
0.017 | - |
fumarate | mutant H505Y, pH 7.5, 25°C | Shewanella frigidimarina | |
0.021 | - |
fumarate | mutant H505Y, pH 9.0, 25°C | Shewanella frigidimarina | |
0.022 | - |
fumarate | mutant H505Y, pH 6.0, 25°C | Shewanella frigidimarina | |
0.025 | - |
fumarate | wild-type enzyme and mutant H505Y, pH 7.2, 25°C | Shewanella frigidimarina | |
0.028 | - |
fumarate | wild-type enzyme, pH 7.5, 25°C | Shewanella frigidimarina | |
0.043 | - |
fumarate | wild-type enzyme and mutant H505A, pH 6.0, 25°C | Shewanella frigidimarina | |
0.109 | - |
fumarate | mutant H505A, pH 7.2, 25°C | Shewanella frigidimarina | |
0.129 | - |
fumarate | mutant H505A, pH 7.5, 25°C | Shewanella frigidimarina |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
soluble | - |
Shewanella frigidimarina | - |
- |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Na+ | internally bound, coordinated by 1 solvent water molecule and the backbone carbonyl oxygens of residues Thr506, Met507, Gly508, Glu534, and Thr536, structure analysis, His505 is not essential for maintaining the sodium binding site | Shewanella frigidimarina |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Shewanella frigidimarina | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant flavocytochrome enzymes from the enzyme-deficient strain DELTAfccA | Shewanella frigidimarina |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
succinate + NAD+ = fumarate + NADH + H+ | His505 influences the active site | Shewanella frigidimarina |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
fumarate + NADH + H+ | - |
Shewanella frigidimarina | succinate + NAD+ | - |
? |
Synonyms | Comment | Organism |
---|---|---|
fumarate reductase | - |
Shewanella frigidimarina |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Shewanella frigidimarina |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
32 | - |
fumarate | mutant H505A, pH 6.0, 25°C | Shewanella frigidimarina | |
79 | - |
fumarate | mutant H505A, pH 7.2, 25°C | Shewanella frigidimarina | |
101 | - |
fumarate | mutant H505A, pH 7.5, 25°C | Shewanella frigidimarina | |
105 | - |
fumarate | mutant H505A, pH 9.0, 25°C | Shewanella frigidimarina | |
210 | - |
fumarate | wild-type enzyme, pH 9.0, 25°C | Shewanella frigidimarina | |
240 | - |
fumarate | mutant H505Y, pH 9.0, 25°C | Shewanella frigidimarina | |
354 | - |
fumarate | mutant H505Y, pH 7.2, 25°C | Shewanella frigidimarina | |
363 | - |
fumarate | mutant H505Y, pH 7.5, 25°C | Shewanella frigidimarina | |
370 | - |
fumarate | wild-type enzyme, pH 7.5, 25°C | Shewanella frigidimarina | |
377 | - |
fumarate | mutant H505Y, pH 6.0, 25°C | Shewanella frigidimarina | |
509 | - |
fumarate | wild-type enzyme, pH 7.2, 25°C | Shewanella frigidimarina | |
658 | - |
fumarate | wild-type enzyme, pH 6.0, 25°C | Shewanella frigidimarina |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Shewanella frigidimarina |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6 | 9 | - |
Shewanella frigidimarina |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
flavocytochrome c3 | - |
Shewanella frigidimarina | |
NADH | - |
Shewanella frigidimarina |