Cloned (Comment) | Organism |
---|---|
cloning and expression of wild-type DDC and Y332F and T246A mutants in SVS370 Escherichia coli cells. | Sus scrofa |
Protein Variants | Comment | Organism |
---|---|---|
T246A | T246 act as an essential general base for the oxidative deamination reaction | Sus scrofa |
Y332F | wild-type enzyme and Y332F variant are able to perform the oxidation toward aromatic amines or aromatic L-amino acids, without the aid of any cofactor related to oxygen chemistry. | Sus scrofa |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
3-(3,4-dihydroxyphenyl)-2-hydrazino-2-methyl propionic acid | carbiDOPA, addition of 10 microM inhibitor to reaction mixtures (Y332F mutant with L-dopa) in the presence or in the absence of catalase or superoxide dismutase, immediately stops the O2 consumption. | Sus scrofa | |
5-hydroxy-L-tryptophan | substrate inhibition | Sus scrofa | |
L-Dopa | substrate inhibition | Sus scrofa |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.049 | - |
5-hydroxy-L-tryptophan | wild-type enzyme | Sus scrofa | |
0.058 | - |
L-Dopa | wild-type enzyme | Sus scrofa |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-hydroxy-L-tryptophan | Sus scrofa | - |
serotonin + CO2 | - |
? | |
L-Dopa | Sus scrofa | - |
dopamine + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Sus scrofa | P80041 | - |
- |
Purification (Comment) | Organism |
---|---|
commentary | Sus scrofa |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
The course of the reaction of Y332 mutant with 0.3 mM L-dopa has been followed at different enzyme concentrations. Although the shapes of the profiles for the interconversion of the 2 coenzymatic forms (pyridoxal 5'-phosphate and pyridoxamine 5'-phosphate) are similar at all the enzyme concentrations tested. At each enzyme concentration there are 2 phases: at first, pyridoxamine 5'-phosphate increases at the expense of pyridoxal 5'-phosphate, and then this tendency is reversed until approximately 85% of the pyridoxal 5'-phosphate cofactor is regenerated, the amount of pyridoxamine 5'-phosphate formed relative to the initial pyridoxal 5'-phosphate content of the enzyme, during consumption of L-dopa, increases as the enzyme concentration decreases. At higher enzyme concentrations, where L-dopa consumption is faster, the accumulation of the pyridoxamine 5'-phosphate-intermediate is less appreciable. In every case the pyridoxamine 5'-phosphate species is generated during the course of linear formation of products. | Sus scrofa |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
5-hydroxy-L-tryptophan | - |
Sus scrofa | serotonin + CO2 | - |
? | |
L-Dopa | - |
Sus scrofa | dopamine + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
DDC | - |
Sus scrofa |
DOPA decarboxylase | - |
Sus scrofa |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | mutant T246A presents a decarboxylase activity, the kcat value is decreased by 29fold with respect to wild-type | Sus scrofa | |
additional information | - |
additional information | When 50 microM Y332F DDC is mixed with 0.1 mM L-dopa under single turnover conditions up to 4 sec a gradual conversion of the pyridoxal 5'-phosphate of the enzyme takes place concomitantly to pyridoxamine 5'-phosphate and aldehyde formation while L-dopa is consumed. After 3.6 sec, 23% of the original pyridoxal 5'-phosphate is converted into pyridoxamine 5'-phosphate and 25% of the original L-dopa is transformed into 3,4-dihydroxyphenylacetaldehyde. A pyridoxamine 5'-phosphate intermediate forms during the first catalytic cycle. | Sus scrofa | |
0.67 | - |
5-hydroxy-L-tryptophan | wild-type enzyme | Sus scrofa | |
4.5 | - |
L-Dopa | Y332F DDC mutant, reaction in 50 mM Hepes, pH 7.5, at 25°C causes the production of ammonia and 3,4-dihydroxyphenylacetaldehyde along with the consumption of molecular oxygen in a 1:2 molar ratio | Sus scrofa | |
8.8 | - |
L-Dopa | wild-type enzyme | Sus scrofa |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Sus scrofa |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
5.7 | - |
5-hydroxy-L-tryptophan | wild-type enzyme, the data of initial velocity for decarboxylation of 5-hydroxy-L-tryptophan versus substrate concentration exhibit a substrate-inhibition pattern that requires a modified version of the Michaelis-Menten equation. | Sus scrofa | |
10 | - |
L-Dopa | wild-type enzyme, the data of initial velocity for decarboxylation of L-dopa versus substrate concentration curiously exhibit a substrate-inhibition pattern that requires a modified version of the Michaelis-Menten equation. | Sus scrofa |