Literature summary extracted from
Carbone, V.; Endo, S.; Sumii, R.; Chung, R.; Matsunaga, T.; Hara, A.; El-Kabbani, O.
Structures of dimeric dihydrodiol dehydrogenase apoenzyme and inhibitor complex: Probing the subunit interface with site-directed mutagenesis (2008), Proteins Struct. Funct. Genet., 70, 176-187.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
1.3.1.20 |
in Escherichia coli BL21(DE3) |
Macaca fascicularis |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
1.3.1.20 |
dimeric apoenzyme (at 2.0 A resolution) and enzyme-inhibitor complex (at 2.9 A resolution), crystals are grown using droplets consisting of the dihydrodiol dehydrogenase/NADPH solution mixed with a matching volume containing 1.5 M ammonium phosphate and 0.1 M sodium citrate at pH 5.6, tertiary structure is formed by a classical dinucleotide binding fold comprising of 2 beta-alpha-beta-alpha-beta motifs at the N-terminus and an eight-stranded, predominantly antiparallel beta-sheet at the C-terminus |
Macaca fascicularis |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
1.3.1.20 |
R148A |
mutagenesis is performed using a QuickChange site-directed mutagenesis kit, dehydrogenase activity is comparable to that of the wild type |
Macaca fascicularis |
1.3.1.20 |
R148A/R202A |
mutagenesis is performed using a QuickChange site-directedmutagenesis kit |
Macaca fascicularis |
1.3.1.20 |
R202A |
mutagenesis is performed using a QuickChange site-directed mutagenesis kit, dehydrogenase activity is comparable to that of the wild type |
Macaca fascicularis |
Inhibitors
EC Number |
Inhibitors |
Comment |
Organism |
Structure |
---|
1.3.1.20 |
4-hydroxyacetophenone |
- |
Macaca fascicularis |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
1.3.1.20 |
70000 |
- |
determined by gel filtration |
Macaca fascicularis |
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
1.3.1.20 |
additional information |
Macaca fascicularis |
dimeric dihydrodiol dehydrogenase catalyses the nicotinamide adenine dinucleotide phosphate NADP+-dependent oxidation of trans-dihydrodiols of aromatic hydrocarbons to their corresponding catechols, dimeric dihydrodiol dehydrogenase is also competent in the NADPH-driven reduction of a limited number of aldehyde and dicarbonyl compounds such as 3-deoxyglucosone, camphorquinone, and 3-nitrobenzaldehyde |
? |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
1.3.1.20 |
Macaca fascicularis |
Q9TQS6 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
1.3.1.20 |
recombinant enzymes are purified from the 12000g supernatants of the homogenates of the cells, the single mutant enzymes are partially purified by the ammonium sulphate fractionation and chromatography on a Sephadex G-100 column, the wild type and double mutant enzyme are further purified by chromatography on a Red Sepharose column |
Macaca fascicularis |
Specific Activity [micromol/min/mg]
EC Number |
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
---|
1.3.1.20 |
0.04 |
- |
R148A and R202A mutant, 25 mM Tris-HCl, pH 7.5, 0.25 mM NADP+ |
Macaca fascicularis |
1.3.1.20 |
0.05 |
- |
wild type enzyme in dimeric form, 25 mM Tris-HCl, pH 7.5, 0.25 mM NADP+ |
Macaca fascicularis |
1.3.1.20 |
1.2 |
- |
monomeric form |
Macaca fascicularis |
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
1.3.1.20 |
additional information |
dimeric dihydrodiol dehydrogenase catalyses the nicotinamide adenine dinucleotide phosphate NADP+-dependent oxidation of trans-dihydrodiols of aromatic hydrocarbons to their corresponding catechols, dimeric dihydrodiol dehydrogenase is also competent in the NADPH-driven reduction of a limited number of aldehyde and dicarbonyl compounds such as 3-deoxyglucosone, camphorquinone, and 3-nitrobenzaldehyde |
Macaca fascicularis |
? |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
1.3.1.20 |
dimer |
site-directed mutagenesis has demonstrated that the intact dimer is not essential for catalytic activity |
Macaca fascicularis |
1.3.1.20 |
More |
the dissociation of DD into the monomer largely affects the stability rather than catalytic activity |
Macaca fascicularis |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
1.3.1.20 |
dihydrodiol dehydrogenase |
- |
Macaca fascicularis |
Temperature Stability [°C]
EC Number |
Temperature Stability Minimum [°C] |
Temperature Stability Maximum [°C] |
Comment |
Organism |
---|
1.3.1.20 |
37 |
- |
no activity of the 37°C-inactivated R148A/R202A mutant is recovered by the addition of 0.1 mM NADP+ and 40% glycerol |
Macaca fascicularis |
pH Optimum
EC Number |
pH Optimum Minimum |
pH Optimum Maximum |
Comment |
Organism |
---|
1.3.1.20 |
7.5 |
- |
assay at |
Macaca fascicularis |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
1.3.1.20 |
NADP+ |
- |
Macaca fascicularis |
|
1.3.1.20 |
NADPH |
- |
Macaca fascicularis |
|
IC50 Value
EC Number |
IC50 Value |
IC50 Value Maximum |
Comment |
Organism |
Inhibitor |
Structure |
---|
1.3.1.20 |
0.00097 |
- |
the hydroxyl group of 4-hydroxyacetophenone forms hydrogen bond interactions with the side-chain oxygen and the main chain nitrogen of Asp280, the benzene ring and methyl group make a number of hydrophobic contacts with Phe279, Leu158 and Leu177 |
Macaca fascicularis |
4-hydroxyacetophenone |
|