Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.42 extracted from

  • Lee, P.; Colman, R.F.
    Implication by site-directed mutagenesis of Arg314 and Tyr316 in the coenzyme site of pig mitochondrial NADP-dependent isocitrate dehydrogenase (2002), Arch. Biochem. Biophys., 401, 81-90.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli TB1 as maltose binding fusion proteins Sus scrofa

Protein Variants

Protein Variants Comment Organism
K321Q site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
R314Q site-directed mutagenesis, 10fold increased Km for NADP+ compared to the wild-type enzyme Sus scrofa
R323Q site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
Y316F site-directed mutagenesis, kinetics are similar to the wild-type enzyme Sus scrofa
Y316L site-directed mutagenesis, 4fold increased Km for NADP+ compared to the wild-type enzyme Sus scrofa

Inhibitors

Inhibitors Comment Organism Structure
NAD+
-
Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0003
-
Mn2+ pH 7.4, recombinant wild-type enzyme Sus scrofa
0.0004
-
Mn2+ pH 7.4, recombinant mutant Y316F and K323Q Sus scrofa
0.0005
-
Mn2+ pH 7.4, recombinant mutant K321Q Sus scrofa
0.0059
-
isocitrate pH 7.4, recombinant mutant Y316F Sus scrofa
0.0062
-
isocitrate pH 7.4, recombinant mutant K323Q Sus scrofa
0.0074
-
isocitrate pH 7.4, recombinant wild-type enzyme Sus scrofa
0.0077
-
NADP+ pH 7.4, recombinant mutant K323Q Sus scrofa
0.008
-
NADP+ pH 7.4, recombinant mutant Y316F and K321Q Sus scrofa
0.009
-
NADP+ pH 7.4, recombinant wild-type enzyme Sus scrofa
0.01
-
isocitrate pH 7.4, recombinant mutant K321Q Sus scrofa
0.032
-
NADP+ pH 7.4, recombinant mutant Y316L Sus scrofa
0.092
-
NADP+ pH 7.4, recombinant mutant R314Q Sus scrofa

Localization

Localization Comment Organism GeneOntology No. Textmining
mitochondrion
-
Sus scrofa 5739
-

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+
-
Sus scrofa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
82000
-
recombinant mutant Y316F, dynamic light scattering Sus scrofa
86000
-
recombinant wild-type enzyme, dynamic light scattering Sus scrofa
87000
-
recombinant mutants K321Q and R314Q, dynamic light scattering Sus scrofa
97900
-
recombinant mutant Y316L, native PAGE at pH 5.5 Sus scrofa
99800
-
recombinant wild-type enzyme, native PAGE at pH 5.5 Sus scrofa

Organism

Organism UniProt Comment Textmining
Sus scrofa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli, to homogeneity by amylose affinity chromatography, thrombin digestion, and DEAE ion exchange chromatography Sus scrofa

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
7
-
purified recombinant mutant Y316L Sus scrofa
18.1
-
purified recombinant mutant R314Q Sus scrofa
38.6
-
purified recombinant wild-type enzyme Sus scrofa
38.7
-
purified recombinant mutant K323Q Sus scrofa
39.6
-
purified recombinant mutant K321Q Sus scrofa
40.3
-
purified recombinant mutant Y316F Sus scrofa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + NADP+
-
Sus scrofa 2-oxoglutarate + CO2 + NADPH + H+
-
?

Subunits

Subunits Comment Organism
dimer residues Arg314, Tyr316, Lys321, and Arg323 are not involved in subunit interaction Sus scrofa

Synonyms

Synonyms Comment Organism
NADP-dependent isocitrate dehydrogenase
-
Sus scrofa

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.4
-
assay at Sus scrofa

Cofactor

Cofactor Comment Organism Structure
NADP+ Arg314 and the aromatic ring of Tyr316 are important for affinity to the cofactor and catalysis, interaction with 2'-phosphate group of NADP+ Sus scrofa

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
8
-
NAD+ pH 7.4, versus NADPH, recombinant mutant R314Q Sus scrofa
11
-
NAD+ pH 7.4, versus NADPH, recombinant wild-type enzyme and mutant Y316L Sus scrofa
12
-
NAD+ pH 7.4, versus NADPH, recombinant mutant Y316F Sus scrofa