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Literature summary for 1.1.1.45 extracted from

  • Ishikura, S.; Usami, N.; Araki, M.; Hara, A.
    Structural and functional characterization of rabbit and human L-gulonate 3-dehydrogenase (2005), J. Biochem., 137, 303-314.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli Oryctolagus cuniculus

Protein Variants

Protein Variants Comment Organism
D36R mutation leads to a switch in the favor of NADP(H) specificity, suggesting an important role of Asp36 in the coenzyme specificity Oryctolagus cuniculus
E157Q mutation produces no large kinetic alterations Oryctolagus cuniculus
H145Q inactive enzyme form Oryctolagus cuniculus
N195D inactive enzyme form Oryctolagus cuniculus
N196Q inactive enzyme form Oryctolagus cuniculus
S124A mutation decreases the catalytic efficiency 500 fold Oryctolagus cuniculus

Inhibitors

Inhibitors Comment Organism Structure
Cibacron blue competitive inhibition with respect to NAD+ Oryctolagus cuniculus
malonate reversible inhibition, IC50: 0.34 mM Oryctolagus cuniculus
malonic acid monoethyl ester weak inhibition, IC50: 5.5 mM Oryctolagus cuniculus
additional information no inhibition by 20 mM D-glucose, 2 mM 2-keto-L-gulonate, 1 mM D-glucorunate or 6 mM ADP, ATP and AMP; no significant inhibition (less than 15%) by 1 mM L-ascorbate, monocarboxylic acids, or dicarboxylic acids Oryctolagus cuniculus
p-chloromercuriphenylsulfonate 0.001 mM, complete inhibition Oryctolagus cuniculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information no activity with 5-40 mM sugars or hydroxyacids like 2-keto-L-guluonate, 3-deoxyglucosone, xylitol, L-threitol, ascorbic acid, malic acid, L-threonine, L-glycerate, L-tartarate and L-lactate, or 50 mM ethyl and methyl esters of 3-hydroxybutyrate Oryctolagus cuniculus
0.0005
-
NADH at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
0.0006
-
NADH at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
0.01
-
NAD+ at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
0.011
-
NAD+ at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
0.18
-
L-gulonate at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
0.21
-
L-gulonate at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
0.49
-
L-gulonate mutant enzyme E157Q, NAD+-linked activity Oryctolagus cuniculus
0.67
-
NADP+ at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
0.89
-
NADP+ at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
1.2
-
L-threonate at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
2.2
-
L-3-hydroxybutyrate at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
2.3
-
L-3-hydroxybutyrate at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
2.9
-
L-threonate at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
4.4
-
L-gulonate mutant enzyme S124A, NAD+-linked activity Oryctolagus cuniculus
8.3
-
acetoacetate at pH 8.5, rabbit liver GDH Oryctolagus cuniculus
9.9
-
acetoacetate at pH 8.5, recombinant rabbit GDH Oryctolagus cuniculus
14
-
L-gulonate mutant enzyme D36R, NAD+-linked activity Oryctolagus cuniculus
18.5
-
L-gulonate mutant enzyme D36R, NADP+-linked activity Oryctolagus cuniculus
18.5
-
L-gulonate mutant enzyme E157Q, NADP+-linked activity Oryctolagus cuniculus

Localization

Localization Comment Organism GeneOntology No. Textmining
cytosol
-
Oryctolagus cuniculus 5829
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
35000
-
2 * 35000 Oryctolagus cuniculus
36000
-
SDS-PAGE Oryctolagus cuniculus
70000
-
gel filtration Oryctolagus cuniculus

Organism

Organism UniProt Comment Textmining
Oryctolagus cuniculus
-
Japanese white rabbit
-

Purification (Commentary)

Purification (Comment) Organism
ammonium sulfate fractionation, Sephadex G-100 column chromatography, Q-Sepharose column chromatography, Blue-Sepharose column chromatography, hydroxylapatite column chromatography Oryctolagus cuniculus

Source Tissue

Source Tissue Comment Organism Textmining
lens
-
Oryctolagus cuniculus
-
liver
-
Oryctolagus cuniculus
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.011
-
in the supernatant Oryctolagus cuniculus
6.71
-
after purification Oryctolagus cuniculus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
? + NADP+
-
Oryctolagus cuniculus ? + NADPH
-
?
acetoacetate + NADH
-
Oryctolagus cuniculus ? + NAD+
-
?
L-3-hydroxybutyrate + ?
-
Oryctolagus cuniculus ?
-
?
L-gulonate + NAD+
-
Oryctolagus cuniculus 3-dehydro-L-gulonate + NADH + H+
-
?
L-threonate + ?
-
Oryctolagus cuniculus ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 35000 Oryctolagus cuniculus

Synonyms

Synonyms Comment Organism
GDH
-
Oryctolagus cuniculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Oryctolagus cuniculus

Cofactor

Cofactor Comment Organism Structure
NADH
-
Oryctolagus cuniculus
NADP+
-
Oryctolagus cuniculus

pI Value

Organism Comment pI Value Maximum pI Value
Oryctolagus cuniculus
-
-
7.3

IC50 Value

IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
0.34
-
reversible inhibition, IC50: 0.34 mM Oryctolagus cuniculus malonate
5.5
-
weak inhibition, IC50: 5.5 mM Oryctolagus cuniculus malonic acid monoethyl ester