Literature summary for 1.17.1.1 extracted from
Gonzalez-Porque, P.
Cytidine diphosphate 4-keto-6-deoxy-D-glucose-3-dehydrogenase (1986), Coenzymes and cofactors, Glutathione, Chem. Biochem. Med. Aspects Pt. A (Dolphin D, Poulson R, Avromonic O, eds. ) John Wiley & Sons, New York, 1, 391-419.
No PubMed abstract available
Inhibitors
Inhibitors |
Comment |
Organism |
Structure |
5,5'-dithiobis(2-nitrobenzoate) |
inhibits the activity of the whole system |
Yersinia pseudotuberculosis |
|
N-ethylmaleimide |
inhibits the activity of the whole system |
Yersinia pseudotuberculosis |
|
Molecular Weight [Da]
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
additional information |
- |
cofactor: 200-400, gel filtration on Bio Gel P2 |
Yersinia pseudotuberculosis |
35000 |
45000 |
enzyme E3, thin-layer chromatography on Sephadex G-100 |
Yersinia pseudotuberculosis |
40000 |
- |
enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone |
Yersinia pseudotuberculosis |
60000 |
70000 |
enzyme E1, thin-layer chromatography on Sephadex G-100 |
Yersinia pseudotuberculosis |
61000 |
- |
enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone |
Yersinia pseudotuberculosis |
Natural Substrates/ Products (Substrates)
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H |
Yersinia pseudotuberculosis |
biosynthesis of 3,6-dideoxysugars |
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O |
- |
? |
|
Organism
Organism |
UniProt |
Comment |
Textmining |
Yersinia pseudotuberculosis |
- |
- |
- |
Purification (Commentary)
Purification (Comment) |
Organism |
of cofactor, using streptomycin sulfate precipitation, ammonium sulfate precipitation and dialysis, DEAE-cellulose chromatography, ultrafiltration, Bio Gel P-2 filtration and ion exchange chromatography on Dowex-1 |
Yersinia pseudotuberculosis |
the first three steps are common to the purification of enzyme E1, enzyme E3 and cofactor, their separation can be accomplished after step 4, step 1: preparation of the crude extract, step 2: streptomycin sulfate precipitation, step 3: ammonium sulfate precipitation and dialysis, step 4: DEAE-cellulose chromatography, step 5: purification of enzyme E1, gel filtration on Sephadex G-100, step 6: second DEAE-cellulose chromatography, step 7: preparative polyacrylamide gel electrophoresis, step 5': purification of enzyme E3, gel filtration on Sephadex G-100, step 6': second DEAE-cellulose chromatography, step 7': third DEAE-cellulose chromatography |
Yersinia pseudotuberculosis |
Reaction
Reaction |
Comment |
Organism |
Reaction ID |
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ |
mechanism |
Yersinia pseudotuberculosis |
|
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ |
2 proteins, E1 and E3, are involved but no partial reaction is observed in the presence of either alone |
Yersinia pseudotuberculosis |
|
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O = CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H + H+ |
role of the enzymes E1 and E3, as well as role of the cofactor. Multicomponent system consisting of substrate, NADH or NADPH, enzyme E1: recognition unit of the system by binding to the substrate through the amino group of the coenzyme, enzyme E3: acts as the dehydrogenase of the system, and cofactor: pyridoxamine 5'-phosphate |
Yersinia pseudotuberculosis |
|
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg] |
Specific Activity Maximum [µmol/min/mg] |
Comment |
Organism |
0.11 |
- |
E1 |
Yersinia pseudotuberculosis |
0.32 |
- |
E3 |
Yersinia pseudotuberculosis |
Storage Stability
Storage Stability |
Organism |
frozen, for months without loss of activity, enzyme E1 |
Yersinia pseudotuberculosis |
lyophilized powder: enzyme E3 not stable |
Yersinia pseudotuberculosis |
lyophilized powder: for months, enzyme E1 |
Yersinia pseudotuberculosis |
Substrates and Products (Substrate)
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H |
the substrate undergoes substitution of a hydroxyl group by a hydrogen atom at carbon atom 3 from glucose, it is supposed that the reaction is triggered by an attack from a hydride ion from NAD(P)H to atom carbon 3 |
Yersinia pseudotuberculosis |
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O |
- |
? |
|
CDP-4-dehydro-6-deoxy-D-glucose + NAD(P)H |
biosynthesis of 3,6-dideoxysugars |
Yersinia pseudotuberculosis |
CDP-4-dehydro-3,6-dideoxy-D-glucose + NAD(P)+ + H2O |
- |
? |
|
Subunits
Subunits |
Comment |
Organism |
monomer |
enzyme E3: 1 * 40000, SDS-PAGE, enzyme E1: 1 * 61000, SDS-PAGE, 2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone |
Yersinia pseudotuberculosis |
Synonyms
Synonyms |
Comment |
Organism |
More |
2 proteins E1 and E3 are involved but no partial reaction has been observed in the presence of either alone |
Yersinia pseudotuberculosis |
Cofactor
Cofactor |
Comment |
Organism |
Structure |
additional information |
3.8 SH groups per molecule of enzyme E1, in the presence and absence of SDS |
Yersinia pseudotuberculosis |
|
additional information |
0.37 SH groups per molecule of enzyme E3, in the absence of SDS and 1 SH group per molecule of enzyme E3 in the presence of SDS, the single SH group in enzyme E3 is essential for activity |
Yersinia pseudotuberculosis |
|
additional information |
derivatives of vitamin B6 such as pyridoxine, pyridoxine 5'-phosphate, pyridoxal, pyridoxal 5'-phosphate, pyridoxamine and pyridoxamine 5'-phosphate have stimulatory effect on the reaction, indicating that the amino group and the phosphate group are necessary for the reaction |
Yersinia pseudotuberculosis |
|
NADH |
the second substrate of the reaction, the hydrogen donor |
Yersinia pseudotuberculosis |
|
NADPH |
the second substrate of the reaction, the hydrogen donor |
Yersinia pseudotuberculosis |
|
pyridoxamine 5'-phosphate |
bound to enzyme E1 through an ionic interaction with a positive charge on the surface of the enzyme, the cofactor is needed for the binding of the substrate to the enzyme |
Yersinia pseudotuberculosis |
|