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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.
    View publication on PubMed

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