BRENDA - Enzyme Database

Structural and functional characterization of rabbit and human L-gulonate 3-dehydrogenase

Ishikura, S.; Usami, N.; Araki, M.; Hara, A.; J. Biochem. 137, 303-314 (2005)

Data extracted from this reference:

Cloned(Commentary)
EC Number
Commentary
Organism
1.1.1.45
expressed in Escherichia coli
Oryctolagus cuniculus
Engineering
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.45
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
1.1.1.45
E157Q
mutation produces no large kinetic alterations
Oryctolagus cuniculus
1.1.1.45
H145Q
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
N195D
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
N196Q
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
S124A
mutation decreases the catalytic efficiency 500 fold
Oryctolagus cuniculus
Inhibitors
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.45
Cibacron blue
competitive inhibition with respect to NAD+
Oryctolagus cuniculus
1.1.1.45
malonate
reversible inhibition, IC50: 0.34 mM
Oryctolagus cuniculus
1.1.1.45
malonic acid monoethyl ester
weak inhibition, IC50: 5.5 mM
Oryctolagus cuniculus
1.1.1.45
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
1.1.1.45
p-chloromercuriphenylsulfonate
0.001 mM, complete inhibition
Oryctolagus cuniculus
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.45
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
1.1.1.45
0.0005
-
NADH
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.0006
-
NADH
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.01
-
NAD+
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.011
-
NAD+
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.18
-
L-gulonate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.21
-
L-gulonate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.49
-
L-gulonate
mutant enzyme E157Q, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
0.67
-
NADP+
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.89
-
NADP+
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
1.2
-
L-threonate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
2.2
-
L-3-hydroxybutyrate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
2.3
-
L-3-hydroxybutyrate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
2.9
-
L-threonate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
4.4
-
L-gulonate
mutant enzyme S124A, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
8.3
-
acetoacetate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
9.9
-
acetoacetate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
14
-
L-gulonate
mutant enzyme D36R, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
18.5
-
L-gulonate
mutant enzyme D36R, NADP+-linked activity; mutant enzyme E157Q, NADP+-linked activity
Oryctolagus cuniculus
Localization
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.45
cytosol
-
Oryctolagus cuniculus
5829
-
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.45
35000
-
2 * 35000
Oryctolagus cuniculus
1.1.1.45
36000
-
SDS-PAGE
Oryctolagus cuniculus
1.1.1.45
70000
-
gel filtration
Oryctolagus cuniculus
Organism
EC Number
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
1.1.1.45
Oryctolagus cuniculus
-
Japanese white rabbit
-
Purification (Commentary)
EC Number
Commentary
Organism
1.1.1.45
ammonium sulfate fractionation, Sephadex G-100 column chromatography, Q-Sepharose column chromatography, Blue-Sepharose column chromatography, hydroxylapatite column chromatography
Oryctolagus cuniculus
Source Tissue
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.45
lens
-
Oryctolagus cuniculus
-
1.1.1.45
liver
-
Oryctolagus cuniculus
-
Specific Activity [micromol/min/mg]
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.45
0.011
-
in the supernatant
Oryctolagus cuniculus
1.1.1.45
6.71
-
after purification
Oryctolagus cuniculus
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.45
? + NADP+
-
669147
Oryctolagus cuniculus
? + NADPH
-
-
-
?
1.1.1.45
acetoacetate + NADH
-
669147
Oryctolagus cuniculus
? + NAD+
-
-
-
?
1.1.1.45
L-3-hydroxybutyrate + ?
-
669147
Oryctolagus cuniculus
?
-
-
-
?
1.1.1.45
L-gulonate + NAD+
-
669147
Oryctolagus cuniculus
3-dehydro-L-gulonate + NADH + H+
-
-
-
?
1.1.1.45
L-threonate + ?
-
669147
Oryctolagus cuniculus
?
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.1.1.45
dimer
2 * 35000
Oryctolagus cuniculus
pH Optimum
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.45
6.5
-
-
Oryctolagus cuniculus
Cofactor
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.45
NADH
-
Oryctolagus cuniculus
1.1.1.45
NADP+
-
Oryctolagus cuniculus
pI Value
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.45
Oryctolagus cuniculus
-
-
7.3
IC50 Value
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
1.1.1.45
0.34
-
reversible inhibition, IC50: 0.34 mM
Oryctolagus cuniculus
malonate
1.1.1.45
5.5
-
weak inhibition, IC50: 5.5 mM
Oryctolagus cuniculus
malonic acid monoethyl ester
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.45
expressed in Escherichia coli
Oryctolagus cuniculus
Cofactor (protein specific)
EC Number
Cofactor
Commentary
Organism
Structure
1.1.1.45
NADH
-
Oryctolagus cuniculus
1.1.1.45
NADP+
-
Oryctolagus cuniculus
Engineering (protein specific)
EC Number
Amino acid exchange
Commentary
Organism
1.1.1.45
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
1.1.1.45
E157Q
mutation produces no large kinetic alterations
Oryctolagus cuniculus
1.1.1.45
H145Q
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
N195D
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
N196Q
inactive enzyme form
Oryctolagus cuniculus
1.1.1.45
S124A
mutation decreases the catalytic efficiency 500 fold
Oryctolagus cuniculus
IC50 Value (protein specific)
EC Number
IC50 Value
IC50 Value Maximum
Commentary
Organism
Inhibitor
Structure
1.1.1.45
0.34
-
reversible inhibition, IC50: 0.34 mM
Oryctolagus cuniculus
malonate
1.1.1.45
5.5
-
weak inhibition, IC50: 5.5 mM
Oryctolagus cuniculus
malonic acid monoethyl ester
Inhibitors (protein specific)
EC Number
Inhibitors
Commentary
Organism
Structure
1.1.1.45
Cibacron blue
competitive inhibition with respect to NAD+
Oryctolagus cuniculus
1.1.1.45
malonate
reversible inhibition, IC50: 0.34 mM
Oryctolagus cuniculus
1.1.1.45
malonic acid monoethyl ester
weak inhibition, IC50: 5.5 mM
Oryctolagus cuniculus
1.1.1.45
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
1.1.1.45
p-chloromercuriphenylsulfonate
0.001 mM, complete inhibition
Oryctolagus cuniculus
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.1.1.45
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
1.1.1.45
0.0005
-
NADH
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.0006
-
NADH
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.01
-
NAD+
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.011
-
NAD+
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.18
-
L-gulonate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.21
-
L-gulonate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
0.49
-
L-gulonate
mutant enzyme E157Q, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
0.67
-
NADP+
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
0.89
-
NADP+
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
1.2
-
L-threonate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
2.2
-
L-3-hydroxybutyrate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
2.3
-
L-3-hydroxybutyrate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
2.9
-
L-threonate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
4.4
-
L-gulonate
mutant enzyme S124A, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
8.3
-
acetoacetate
at pH 8.5, rabbit liver GDH
Oryctolagus cuniculus
1.1.1.45
9.9
-
acetoacetate
at pH 8.5, recombinant rabbit GDH
Oryctolagus cuniculus
1.1.1.45
14
-
L-gulonate
mutant enzyme D36R, NAD+-linked activity
Oryctolagus cuniculus
1.1.1.45
18.5
-
L-gulonate
mutant enzyme D36R, NADP+-linked activity; mutant enzyme E157Q, NADP+-linked activity
Oryctolagus cuniculus
Localization (protein specific)
EC Number
Localization
Commentary
Organism
GeneOntology No.
Textmining
1.1.1.45
cytosol
-
Oryctolagus cuniculus
5829
-
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.1.1.45
35000
-
2 * 35000
Oryctolagus cuniculus
1.1.1.45
36000
-
SDS-PAGE
Oryctolagus cuniculus
1.1.1.45
70000
-
gel filtration
Oryctolagus cuniculus
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
1.1.1.45
ammonium sulfate fractionation, Sephadex G-100 column chromatography, Q-Sepharose column chromatography, Blue-Sepharose column chromatography, hydroxylapatite column chromatography
Oryctolagus cuniculus
Source Tissue (protein specific)
EC Number
Source Tissue
Commentary
Organism
Textmining
1.1.1.45
lens
-
Oryctolagus cuniculus
-
1.1.1.45
liver
-
Oryctolagus cuniculus
-
Specific Activity [micromol/min/mg] (protein specific)
EC Number
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.1.1.45
0.011
-
in the supernatant
Oryctolagus cuniculus
1.1.1.45
6.71
-
after purification
Oryctolagus cuniculus
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
1.1.1.45
? + NADP+
-
669147
Oryctolagus cuniculus
? + NADPH
-
-
-
?
1.1.1.45
acetoacetate + NADH
-
669147
Oryctolagus cuniculus
? + NAD+
-
-
-
?
1.1.1.45
L-3-hydroxybutyrate + ?
-
669147
Oryctolagus cuniculus
?
-
-
-
?
1.1.1.45
L-gulonate + NAD+
-
669147
Oryctolagus cuniculus
3-dehydro-L-gulonate + NADH + H+
-
-
-
?
1.1.1.45
L-threonate + ?
-
669147
Oryctolagus cuniculus
?
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.1.1.45
dimer
2 * 35000
Oryctolagus cuniculus
pH Optimum (protein specific)
EC Number
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
1.1.1.45
6.5
-
-
Oryctolagus cuniculus
pI Value (protein specific)
EC Number
Organism
Commentary
pI Value Maximum
pI Value
1.1.1.45
Oryctolagus cuniculus
-
-
7.3