Cloned (Comment) | Organism |
---|---|
gene A3L14_07690, recombinant enzyme expression in Escherichia coli strain Rosetta2 | Thermococcus thioreducens |
Crystallization (Comment) | Organism |
---|---|
purified enzyme in unit cells belonging to space groups P21, P212121 and P43212 (monoclinic, orthorhombic, and tetragonal crystals), vapour diffusion method, mixing of 0.0012 ml of 10 mg/ml protein in 50 mM Tris, pH 7.5, and 50 mM NaCl, with 600 nl of reservoir solution containing 8% PEG 4000, pH 4.2-4.5, in a protein:reservoir ratio of 2:1 (1:1 for the tetragonal crystals), at 20°C, X-ray diffraction structure determination and analysis at 2.4 A, 2.1 A, and 1.9 A resolution, respectively, modeling by molecular replacement using the FeADH from Thermotoga maritima as a template | Thermococcus thioreducens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | required, the iron is internally sequestered, and is bound entirely by amino acids from one domain: three histidines and one aspartic acid. The monoclinic and orthorhombic asymmetric units one molecule contained iron and an NADP molecule, while the other do not. The tetragonal crystals lack both iron and NADP+ | Thermococcus thioreducens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus thioreducens | A0A0Q2QQL1 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant enzyme from Escherichia coli strain Rosetta2 by heat treatment for 30 min at 75°C, the supernatant is further purified by cation exchange and anion exchange chromatography, ultrafiltration, and gel filtration | Thermococcus thioreducens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
1,4-butanediol + NADP+ | - |
Thermococcus thioreducens | 4-hydroxybutanal + NADPH + H+ | - |
r |
Subunits | Comment | Organism |
---|---|---|
dimer | x * 41500, SDS-PAGE | Thermococcus thioreducens |
Synonyms | Comment | Organism |
---|---|---|
A3L14_07690 | - |
Thermococcus thioreducens |
FeADH | - |
Thermococcus thioreducens |
iron-containing alcohol dehydrogenase | - |
Thermococcus thioreducens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | the coenzyme is in an extended conformation, a feature that is common to the large superfamily of NADH-dependent dehydrogenases that share a classical nucleotide-binding domain. A long broad tunnel passes entirely through the enzyme between the two domains, completely encapsulating the coenzyme | Thermococcus thioreducens | |
NADP+ | - |
Thermococcus thioreducens | |
NADPH | - |
Thermococcus thioreducens |