Cloned (Comment) | Organism |
---|---|
expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Gold | Aspergillus nidulans |
Crystallization (Comment) | Organism |
---|---|
purified recombinant His-tagged enzyme, microbatch method, 4°C, mixing of equal volumes of protein and precipitant solution, the latter containing 20% w/v PEG 3350, 0.1 M Bis-Tris propane pH 7.5, 0.2 M sodium fluoride, cryoprotection with 25% glycerol, X-ray diffraction structure determination and analysis at 2.8 A resolution, molecular replacement | Aspergillus nidulans |
Protein Variants | Comment | Organism |
---|---|---|
K160V | site-directed mutagenesis of the key residue responsible for anchoring of the co-factor, inactive mutant | Aspergillus nidulans |
L320C | site-directed mutagenesis of a gate-keeper residue, the L320C mutant enzyme is also active in UDPGlcpNAc/UDP-GalpNAc interconversion | Aspergillus nidulans |
L320Y | site-directed mutagenesis of a gate-keeper residue, the L320Y mutant enzyme is not active in UDPGlcpNAc/UDP-GalpNAc interconversion | Aspergillus nidulans |
Y156F | site-directed mutagenesis of the key residue serving as the active site base, inactive mutant | Aspergillus nidulans |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
UDP-N-acetylgalactosamine | competitive, binding study | Aspergillus nidulans | |
UDP-N-acetylglucosamine | competitive, binding study | Aspergillus nidulans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetic analyis of wild-type and mutant enzymes, overview | Aspergillus nidulans | |
0.06 | - |
UDP-alpha-D-glucose | mutant L320Y, pH 8.5, 25°C | Aspergillus nidulans | |
0.1 | 1 | UDP-alpha-D-glucose | wild-type enzyme, pH 8.5, 25°C | Aspergillus nidulans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40600 | - |
x * 40600 | Aspergillus nidulans |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-glucose | Aspergillus nidulans | via 4-ketose intermediate | UDP-alpha-D-galactose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aspergillus nidulans | C8VAU8 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Gold by nickel affinity chromatography and ultrafiltration | Aspergillus nidulans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-alpha-D-glucose | via 4-ketose intermediate | Aspergillus nidulans | UDP-alpha-D-galactose | - |
r | |
UDP-alpha-D-glucose | via 4-ketose intermediate, UDP-glucose is bound within the active site cleft, substrate binding structure, overview | Aspergillus nidulans | UDP-alpha-D-galactose | - |
r |
Subunits | Comment | Organism |
---|---|---|
? | x * 40600 | Aspergillus nidulans |
Synonyms | Comment | Organism |
---|---|---|
GalE | - |
Aspergillus nidulans |
UDP-galactose-4-epimerase | - |
Aspergillus nidulans |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Aspergillus nidulans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
UDP-alpha-D-glucose | mutant L320Y, pH 8.5, 25°C | Aspergillus nidulans | |
12.8 | - |
UDP-alpha-D-glucose | wild-type enzyme, pH 8.5, 25°C | Aspergillus nidulans |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Aspergillus nidulans |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
NAD+ | bound within the active site cleft, binding structure, overview | Aspergillus nidulans |
General Information | Comment | Organism |
---|---|---|
additional information | ligand binding structures, docking study, overview | Aspergillus nidulans |
physiological function | the enzyme produces the precursor UDP-galactopyranose required for galactofuranose synthesis | Aspergillus nidulans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
26.8 | - |
UDP-alpha-D-glucose | mutant L320Y, pH 8.5, 25°C | Aspergillus nidulans | |
116.4 | - |
UDP-alpha-D-glucose | wild-type enzyme, pH 8.5, 25°C | Aspergillus nidulans |