BRENDA - Enzyme Database
show all sequences of 2.7.7.69

A second GDP-L-galactose phosphorylase in Arabidopsis en route to vitamin C. Covalent intermediate and substrate requirements for the conserved reaction

Linster, C.L.; Adler, L.N.; Webb, K.; Christensen, K.C.; Brenner, C.; Clarke, S.G.; J. Biol. Chem. 283, 18483-18492 (2008)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
-
Arabidopsis thaliana
Engineering
Amino acid exchange
Commentary
Organism
G224D
less than 0.1% of wild-type activity
Arabidopsis thaliana
H238N
less than 0.1% of wild-type activity
Arabidopsis thaliana
S290F
less than 0.1% of wild-type activity
Arabidopsis thaliana
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0044
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
0.0079
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
0.0083
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
0.012
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
0.22
-
phosphate
presence of GDP-G-glucose, pH 7.5, 26°C
Arabidopsis thaliana
0.52
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
0.76
-
phosphate
presence of GDP-alpha-D-glucose, pH 7.5, 26°C
Arabidopsis thaliana
1
-
phosphate
presence of GDP-L-galactose, pH 7.5, 26°C
Arabidopsis thaliana
1.3
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
2.4
-
phosphate
presence of GDP-alpha-L-galactose, pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-L-galactose 1-phosphate
pH 7.5, 26°C; pH 7.5, 26°C
Arabidopsis thaliana
10
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
16
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
29
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
45
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
GDP-beta-L-galactose + phosphate
Arabidopsis thaliana
enzyme guanylylates a conserved active site His residue with GDP-L-galactose, forming L-galactose 1-phosphate for vitamin C synthesis, and regeneration of the enzyme with phosphate to form GDP
GDP + beta-L-galactose 1-phosphate
-
-
r
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Arabidopsis thaliana
Q8RWE8
isoform Vtc2
-
Arabidopsis thaliana
Q9FLP9
isoform Vtc5
-
Specific Activity [micromol/min/mg]
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
14.2
-
pH 7.5, 26°C, substrate GDP-alpha-D-glucose
Arabidopsis thaliana
15.1
-
pH 7.5, 26°C, substrate GDP-alpha-L-galactose
Arabidopsis thaliana
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
alpha-D-glucose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-D-glucose + phosphate
-
-
-
r
alpha-D-mannose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-D-mannose + phosphate
-
-
-
r
alpha-L-galactose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-L-galactose + phosphate
-
-
-
r
GDP-alpha-D-galactose + phosphate
-
693081
Arabidopsis thaliana
GDP + alpha-D-galactose 1-phosphate
-
-
-
r
GDP-alpha-D-glucose + phosphate
-
693081
Arabidopsis thaliana
alpha-D-glucose 1-phosphate + GDP
-
-
-
r
GDP-alpha-D-mannose + phosphate
-
693081
Arabidopsis thaliana
alpha-D-mannose 1-phosphate + GDP
-
-
-
r
GDP-beta-L-galactose + phosphate
enzyme guanylylates a conserved active site His residue with GDP-L-galactose, forming L-galactose 1-phosphate for vitamin C synthesis, and regeneration of the enzyme with phosphate to form GDP
693081
Arabidopsis thaliana
GDP + beta-L-galactose 1-phosphate
-
-
-
r
GDP-beta-L-galactose + phosphate
reaction proceeds through a covalent guanylated histidine residue within the histidine triad motif
693081
Arabidopsis thaliana
GDP + beta-L-galactose 1-phosphate
-
-
-
r
additional information
poor reactivity of hexose 1-phosphates as acceptors
693081
Arabidopsis thaliana
?
-
-
-
-
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.024
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
0.093
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
3.8
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
5.8
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-L-galactose 1-phosphate
pH 7.5, 26°C; pH 7.5, 26°C
Arabidopsis thaliana
9.9
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
12
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
13
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
23
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
27
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
Cloned(Commentary) (protein specific)
Commentary
Organism
-
Arabidopsis thaliana
Engineering (protein specific)
Amino acid exchange
Commentary
Organism
G224D
less than 0.1% of wild-type activity
Arabidopsis thaliana
H238N
less than 0.1% of wild-type activity
Arabidopsis thaliana
S290F
less than 0.1% of wild-type activity
Arabidopsis thaliana
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0044
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
0.0079
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
0.0083
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
0.012
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
0.22
-
phosphate
presence of GDP-G-glucose, pH 7.5, 26°C
Arabidopsis thaliana
0.52
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
0.76
-
phosphate
presence of GDP-alpha-D-glucose, pH 7.5, 26°C
Arabidopsis thaliana
1
-
phosphate
presence of GDP-L-galactose, pH 7.5, 26°C
Arabidopsis thaliana
1.3
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
2.4
-
phosphate
presence of GDP-alpha-L-galactose, pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-L-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
10
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
16
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
29
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
45
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
GDP-beta-L-galactose + phosphate
Arabidopsis thaliana
enzyme guanylylates a conserved active site His residue with GDP-L-galactose, forming L-galactose 1-phosphate for vitamin C synthesis, and regeneration of the enzyme with phosphate to form GDP
GDP + beta-L-galactose 1-phosphate
-
-
r
Specific Activity [micromol/min/mg] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
14.2
-
pH 7.5, 26°C, substrate GDP-alpha-D-glucose
Arabidopsis thaliana
15.1
-
pH 7.5, 26°C, substrate GDP-alpha-L-galactose
Arabidopsis thaliana
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
alpha-D-glucose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-D-glucose + phosphate
-
-
-
r
alpha-D-mannose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-D-mannose + phosphate
-
-
-
r
alpha-L-galactose 1-phosphate + GDP
-
693081
Arabidopsis thaliana
GDP-alpha-L-galactose + phosphate
-
-
-
r
GDP-alpha-D-galactose + phosphate
-
693081
Arabidopsis thaliana
GDP + alpha-D-galactose 1-phosphate
-
-
-
r
GDP-alpha-D-glucose + phosphate
-
693081
Arabidopsis thaliana
alpha-D-glucose 1-phosphate + GDP
-
-
-
r
GDP-alpha-D-mannose + phosphate
-
693081
Arabidopsis thaliana
alpha-D-mannose 1-phosphate + GDP
-
-
-
r
GDP-beta-L-galactose + phosphate
enzyme guanylylates a conserved active site His residue with GDP-L-galactose, forming L-galactose 1-phosphate for vitamin C synthesis, and regeneration of the enzyme with phosphate to form GDP
693081
Arabidopsis thaliana
GDP + beta-L-galactose 1-phosphate
-
-
-
r
GDP-beta-L-galactose + phosphate
reaction proceeds through a covalent guanylated histidine residue within the histidine triad motif
693081
Arabidopsis thaliana
GDP + beta-L-galactose 1-phosphate
-
-
-
r
additional information
poor reactivity of hexose 1-phosphates as acceptors
693081
Arabidopsis thaliana
?
-
-
-
-
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.024
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
0.093
-
GDP-alpha-D-mannose
pH 7.5, 26°C
Arabidopsis thaliana
3.8
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
5.8
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-D-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
6
-
alpha-L-galactose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
9.9
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
12
-
alpha-D-glucose 1-phosphate
pH 7.5, 26°C
Arabidopsis thaliana
13
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
23
-
GDP-alpha-D-glucose
pH 7.5, 26°C
Arabidopsis thaliana
27
-
GDP-alpha-L-galactose
pH 7.5, 26°C
Arabidopsis thaliana
Other publictions for EC 2.7.7.69
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
739282
Wang
Overexpression of tomato GDP-L ...
Solanum lycopersicum
Plant Cell Rep.
33
1441-1451
2014
-
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4
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1
1
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725801
Wang
Antisense-mediated depletion o ...
Solanum lycopersicum
J. Plant Physiol.
170
303-314
2013
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2
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4
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2
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1
1
-
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-
738329
Hou
Expression of a GDP-L-galactos ...
Vitis pseudoreticulata, Vitis pseudoreticulata Baihe-35-l
Genet. Mol. Res.
12
3830-3844
2013
-
-
-
-
-
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2
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5
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5
-
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-
1
1
1
1
-
-
739380
Li
Light and abiotic stresses reg ...
Actinidia deliciosa, Actinidia eriantha
Planta
238
535-547
2013
-
-
-
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5
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2
2
2
2
-
-
693081
Linster
A second GDP-L-galactose phosp ...
Arabidopsis thaliana
J. Biol. Chem.
283
18483-18492
2008
-
-
1
-
3
-
-
15
-
-
-
2
-
8
-
-
-
-
-
-
2
-
17
-
-
-
-
11
-
-
-
-
-
-
-
-
-
1
-
-
3
-
-
-
-
16
-
-
-
2
-
-
-
-
-
-
2
-
17
-
-
-
-
12
-
-
-
-
-
-
-
-
-
-
694703
Mueller-Moule
An expression analysis of the ...
Arabidopsis thaliana
Plant Mol. Biol.
68
31-41
2008
-
-
-
-
-
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2
-
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1
-
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2
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2
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2
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692994
Linster
Arabidopsis VTC2 encodes a GDP ...
Arabidopsis thaliana
J. Biol. Chem.
282
18879-18885
2007
-
-
-
-
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-
7
-
-
-
-
-
4
-
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-
-
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11
-
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-
7
-
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7
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11
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7
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694667
Dowdle
Two genes in Arabidopsis thali ...
Arabidopsis thaliana
Plant J.
52
673-689
2007
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1
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4
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4
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2
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12
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2
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2
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4
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2
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12
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2
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