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

BRENDA support

Literature summary for 1.1.1.42 extracted from

  • Kim, T.K.; Colman, R.F.
    Ser95, Asn97, and Thr78 are important for the catalytic function of porcine NADP-dependent isocitrate dehydrogenase (2005), Protein Sci., 14, 140-147.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 1.1.1.42

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
N97A site-directed mutagenesis, decreased Vmax compared to the wild-type enzyme, slightly affected Km values, but increased pKa of the ionizable metal-liganded hydroxyl of enzyme-bound isocitrate compared to the wild-type enzyme Sus scrofa
N97D site-directed mutagenesis, highly decreased Vmax compared to the wild-type enzyme Sus scrofa
S95A site-directed mutagenesis, decreased Vmax, and increased Km for isocitrate and Mn2+ compared to the wild-type enzyme Sus scrofa
S95D site-directed mutagenesis, highly decreased Vmax compared to the wild-type enzyme Sus scrofa
T78A site-directed mutagenesis, decreased Vmax, and increased Km for isocitrate and Mn2+ compared to the wild-type enzyme Sus scrofa
T78D site-directed mutagenesis, decreased Vmax compared to the wild-type enzyme Sus scrofa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00011
-
 Mn2+ recombinant wild-type enzyme, pH 7.4, 25°C Sus scrofa
0.0004
-
 Mn2+ recombinant mutant N97A, pH 7.4, 25°C Sus scrofa
0.0012
-
 NADP+ recombinant mutant N97D, pH 7.4, 25°C Sus scrofa
0.0024
-
 Mn2+ recombinant mutant N97D, pH 7.4, 25°C Sus scrofa
0.0033
-
 Mn2+ recombinant mutant T78A, pH 7.4, 25°C Sus scrofa
0.0049
-
 NADP+ recombinant mutant N97A, pH 7.4, 25°C Sus scrofa
0.0051
-
 NADP+ recombinant mutant S95A, pH 7.4 Sus scrofa
0.0053
-
 NADP+ recombinant mutant T78A, pH 7.4, 25°C Sus scrofa
0.0075
-
 isocitrate recombinant wild-type enzyme, pH 7.4, 25°C Sus scrofa
0.0099
-
 NADP+ recombinant wild-type enzyme, pH 7.4, 25°C Sus scrofa
0.0108
-
 NADP+ recombinant mutant T78D, pH 7.4, 25°C Sus scrofa
0.0112
-
 Mn2+ recombinant mutant S95A, pH 7.4, 25°C Sus scrofa
0.014
-
 isocitrate recombinant mutant N97A, pH 7.4, 25°C Sus scrofa
0.0237
-
 NADP+ recombinant mutant S95D, pH 7.4, 25°C Sus scrofa
0.0288
-
 Mn2+ recombinant mutant T78D, pH 7.4, 25°C Sus scrofa
0.066
-
 isocitrate recombinant mutant T78A, pH 7.4, 25°C Sus scrofa
0.0668
-
 Mn2+ recombinant mutant S95D, pH 7.4, 25°C Sus scrofa
0.072
-
 isocitrate recombinant mutant N97D, pH 7.4, 25°C Sus scrofa
0.101
-
 isocitrate recombinant mutant S95A, pH 7.4, 25°C Sus scrofa
0.367
-
 isocitrate recombinant mutant T78D, pH 7.4, 25°C Sus scrofa
2.07
-
 isocitrate recombinant mutant S95D, pH 7.4, 25°C Sus scrofa

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ as Mn2+-isocitrate complex, Ser95, Asn97, and Thr78 are involved in binding Sus scrofa

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
87000
-
 recombinant mutant N97A, native PAGE Sus scrofa
94000
-
 recombinant mutant T78A, native PAGE Sus scrofa
97000
-
 recombinant mutant N97D, native PAGE Sus scrofa
98000
-
 recombinant mutant T78D, native PAGE Sus scrofa
101000
-
 recombinant mutant S95A, native PAGE Sus scrofa
104000
-
 recombinant wild-type enzyme and mutant S95D, native PAGE Sus scrofa

Natural Substrates/ Products (Substrates)

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

Organism

Organism UniProt Comment Textmining
Sus scrofa
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli, cleavage of the fusion proteins by thrombin, to homogeneity by amylose affinity chromatography Sus scrofa

Reaction

Reaction Comment Organism Reaction ID
isocitrate + NADP+ = 2-oxoglutarate + CO2 + NADPH + H+ Ser95, Asn97, and Thr78 are important for the catalysis having distinguishable functions, overview Sus scrofa
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.06
-
 purified recombinant mutant S95D Sus scrofa
0.44
-
 purified recombinant mutant N97D Sus scrofa
5.29
-
 purified recombinant mutant N97A Sus scrofa
6.22
-
 purified recombinant mutant T78D Sus scrofa
7.18
-
 purified recombinant mutant S95A Sus scrofa
12.7
-
 purified recombinant mutant T78A Sus scrofa
35.1
-
 purified recombinant wild-type enzyme Sus scrofa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
isocitrate + NADP+
-
 Sus scrofa 2-oxoglutarate + CO2 + NADPH + H+
-
 ?
isocitrate + NADP+ Ser95, Asn97, and Thr78 are involved in substrate binding Sus scrofa 2-oxoglutarate + CO2 + NADPH + H+
-
 ?

Subunits

Subunits Comment Organism
dimer wild-type and mutant enzymes, circular dichroism and native PAGE Sus scrofa

Synonyms

Synonyms Comment Organism
NADP-dependent isocitrate dehydrogenase
-
 Sus scrofa

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Sus scrofa

pH Optimum

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

pH Range

pH Minimum pH Maximum Comment Organism
5 8 pH-profile of wild-type and alanine-substituted mutant enzymes Sus scrofa

Cofactor

Cofactor Comment Organism Structure
NADP+
-
 Sus scrofa