Inhibitors | Comment | Organism | Structure |
---|---|---|---|
5,5'-dithiobis(2-nitrobenzoate) | - |
Yersinia pseudotuberculosis | |
N-ethylmaleimide | enzyme E1: dithiothreitol in excess protects against inhibition | Yersinia pseudotuberculosis |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
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 |
50000 | 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 |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Yersinia pseudotuberculosis | - |
25 VO | - |
Yersinia pseudotuberculosis 25VO | - |
25 VO | - |
Purification (Comment) | Organism |
---|---|
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 | 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+ | E1 binds the substrate pyridoxamine 5'-phosphate essential for binding, E3 possesses NADH oxidase activity, may be the reductase | 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 Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Yersinia pseudotuberculosis |
Storage Stability | Organism |
---|---|
lyophilized powder: enzyme E3 not stable | Yersinia pseudotuberculosis |
lyophilized powder: for months, enzyme E1 | Yersinia pseudotuberculosis |
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 |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Yersinia pseudotuberculosis |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Yersinia pseudotuberculosis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | 4.3 SH per mol of enzyme E1 in the presence of SDS, calculated using a molecular weight of 61000 Da | 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 | |
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 |