Protein Variants | Comment | Organism |
---|---|---|
R181K | site-directed mutagenesis, the mutant shows a 100fold increase in the Km for malate, a 30fold increase in the Ki for oxalate, and a 10fold increase in Ki for NADH, but only a slight or no change in KNAD compared to the wild-type enzyme | Ascaris suum |
R181Q | site-directed mutagenesis, the mutant shows a 100fold increase in the Km for malate, a 30fold increase in the Ki for oxalate, and a 10fold increase in Ki for NADH, but only a slight or no change in KNAD compared to the wild-type enzyme. The activity of the R181Q mutant can be partially rescued by ammonium ion likely by binding in the pocket vacated by the guanidinium group of R181 | Ascaris suum |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
oxalate | very tight binding inhibitor of the NAD-malic enzyme | Ascaris suum |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of wild-type and mutant enzymes, primary deuterium and 13C isotope effects of mutant R181Q in the absence and presence of ammonium ions, overview | Ascaris suum | |
0.035 | - |
NAD+ | pH 8.5, 25°C, recombinant wild-type enzyme | Ascaris suum | |
0.07 | - |
NAD+ | pH 8.5, 25°C, recombinant mutant R181Q | Ascaris suum | |
0.53 | - |
(S)-malate | pH 8.5, 25°C, recombinant wild-type enzyme | Ascaris suum | |
2 | 3 | (S)-malate | pH 8.5, 25°C, recombinant mutant R181Q in presence of 60 mM guanidinium | Ascaris suum | |
8 | - |
(S)-malate | pH 8.5, 25°C, recombinant mutant R181Q, in presence of 60 mM NH4+ | Ascaris suum | |
50 | - |
(S)-malate | pH 8.5, 25°C, recombinant mutant R181Q | Ascaris suum | |
57 | - |
(S)-malate | pH 8.5, 25°C, recombinant mutant R181K | Ascaris suum |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Ascaris suum | |
NH4+ | partially rescues the activity of the R181Q mutant by binding in the pocket vacated by the guanidinium group of R181, 2 mol of ammonia bind per mole of active sites, high-affinity Km is 0.7 mM, low-affinity Km is 420 mM | Ascaris suum |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | Ascaris suum | - |
pyruvate + CO2 + NADH | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Ascaris suum | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-malate + NAD+ | - |
Ascaris suum | pyruvate + CO2 + NADH | - |
r | |
(S)-malate + NAD+ | the NAD-malic enzyme catalyzes the oxidative decarboxylation of (S)-malate via oxaloacetate, Arg181 is within hydrogen bonding distance of the 1-carboxylate of malate in the active site of the enzyme and interacts with the carboxamide side chain of the nicotinamide ring of NADH, but not with NAD+ | Ascaris suum | pyruvate + CO2 + NADH | - |
r |
Synonyms | Comment | Organism |
---|---|---|
NAD-malic enzyme | - |
Ascaris suum |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at, oxidation reaction | Ascaris suum |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at, oxidation reaction | Ascaris suum |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
additional information | Arg181 is within hydrogen bonding distance of the 1-carboxylate of malate in the active site of the enzyme and interacts with the carboxamide side chain of the nicotinamide ring of NADH, but not with NAD+ | Ascaris suum | |
NAD+ | - |
Ascaris suum | |
NADH | - |
Ascaris suum |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | inhibition patterns by measuring the initial rate as a function of malate or NAD+ with NAD+ or malate maintained equal to its Km at different fixed concentrations of oxalate, including zero, Ki, 2Ki, and 4Ki at pH 7.0 and 30 mM free Mg2+ | Ascaris suum | |
0.006 | - |
oxalate | pH 7.0, 25°C, recombinant wild-type enzyme | Ascaris suum |