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Literature summary for 4.1.1.112 extracted from

  • Narayanan, B.C.; Niu, W.; Han, Y.; Zou, J.; Mariano, P.S.; Dunaway-Mariano, D.; Herzberg, O.
    Structure and function of PA4872 from Pseudomonas aeruginosa, a novel class of oxaloacetate decarboxylase from the PEP mutase/isocitrate lyase superfamily (2008), Biochemistry, 47, 167-182.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 4.1.1.112

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 (DE3) cells Pseudomonas aeruginosa

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging drop vapour diffusion method, aliquots of 10 mg/ml protein in 5 mM MgCl2, 5 mM phosphonopyruvate and 10 mM NaHEPES (pH 7.0) mixed with equal volumes of reservoir solution containing 12% polyethylene glycol 20000 and 0.1 M MES (pH 6.0) Pseudomonas aeruginosa

Protein Variants

Protein Variants Comment Organism
H235A the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme Pseudomonas aeruginosa
H235Q the kcat value for catalysis of oxaloacetate decarboxylation is less than an order of magnitude smaller compared to the wild type enzyme Pseudomonas aeruginosa
Y212F 25fold reduction of the Km value compared to the wild type enzyme Pseudomonas aeruginosa

Inhibitors

Inhibitors Comment Organism Structure
3,3-difluoroxaloacetate
-
 Pseudomonas aeruginosa
Acetopyruvate
-
 Pseudomonas aeruginosa
alpha-ketovalerate
-
 Pseudomonas aeruginosa
oxalate
-
 Pseudomonas aeruginosa
Phosphonopyruvate
-
 Pseudomonas aeruginosa
pyruvate
-
 Pseudomonas aeruginosa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.087
-
 oxaloacetate mutant enzyme Y212F, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa
0.63
-
 3-methyloxaloacetate wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa
1.02
-
 oxaloacetate mutant enzyme H235A, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa
2.2
-
 oxaloacetate wild type enzyme, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa
2.5
-
 oxaloacetate mutant enzyme H235Q, in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ cofactor Pseudomonas aeruginosa

Organism

Organism UniProt Comment Textmining
Pseudomonas aeruginosa Q9HUU1
-
-

Purification (Commentary)

Purification (Comment) Organism
DEAE-cellulose column chromatography, phenyl-Sepharose columnn chromatography, and butyl-Sepharose column chromatography Pseudomonas aeruginosa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3-methyloxaloacetate
-
 Pseudomonas aeruginosa 2-oxobutanoate + CO2
-
 ?
Oxaloacetate
-
 Pseudomonas aeruginosa Pyruvate + CO2
-
 r

Subunits

Subunits Comment Organism
dimer dimer of dimers, X-ray crystallography Pseudomonas aeruginosa

Synonyms

Synonyms Comment Organism
PA4872 member of the PEP mutase/isocitrate lyase superfamily Pseudomonas aeruginosa

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
250
-
 3-methyloxaloacetate in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa
7500
-
 oxaloacetate in 0.2 mM NADH, 5 mM MgCl2 and 50 mM K+HEPES (pH 7.5), at 25°C Pseudomonas aeruginosa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.043
-
 oxalate
-
 Pseudomonas aeruginosa
0.45
-
 3,3-difluoroxaloacetate
-
 Pseudomonas aeruginosa
1.09
-
 Acetopyruvate
-
 Pseudomonas aeruginosa
3
-
 Phosphonopyruvate
-
 Pseudomonas aeruginosa
6.7
-
 alpha-ketovalerate
-
 Pseudomonas aeruginosa
7.2
-
 pyruvate
-
 Pseudomonas aeruginosa