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Literature summary for 1.1.1.39 extracted from

  • Karsten, W.E.; Cook, P.F.
    Multiple roles of arginine 181 in binding and catalysis in the NAD-malic enzyme from Ascaris suum (2007), Biochemistry, 46, 14578-14588.
    View publication on PubMed
Search for more literature for 1.1.1.39

Protein Variants

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

Inhibitors Comment Organism Structure
oxalate very tight binding inhibitor of the NAD-malic enzyme Ascaris suum

KM Value [mM]

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

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/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
(S)-malate + NAD+ Ascaris suum
-
 pyruvate + CO2 + NADH
-
 r

Organism

Organism UniProt Comment Textmining
Ascaris suum
-
-
-

Substrates and Products (Substrate)

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

Synonyms Comment Organism
NAD-malic enzyme
-
 Ascaris suum

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at, oxidation reaction Ascaris suum

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at, oxidation reaction Ascaris suum

Cofactor

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 [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