Cloned (Comment) | Organism |
---|---|
overexpression of wild-type and mutants in strain BL21(DE3)pLysS | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
C52S | site-directed mutagenesis, 3000fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level | Escherichia coli |
C55S | site-directed mutagenesis, 600fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level | Escherichia coli |
E182A | site-directed mutagenesis, 50% activity compared to wild-type, normal formation of reaction intermediate UMP-enzyme, contains reduced zinc content and no iron | Escherichia coli |
H115N | site-directed mutagenesis, 2.9% activity compared to wild-type, slightly reduced formation of reaction intermediate UMP-enzyme, retention of zinc and iron | Escherichia coli |
H164N | site-directed mutagenesis, 10000fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1,10-phenanthroline | mutant E182A | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia coli | |
0.048 | - |
UDP-galactose | recombinant mutant E182A, pH 8.5, 27°C | Escherichia coli | |
0.12 | - |
UDP-galactose | recombinant wild-type, pH 8.5, 27°C | Escherichia coli | |
0.16 | - |
alpha-D-glucose 1-phosphate | recombinant wild-type, pH 8.5, 27°C | Escherichia coli | |
0.19 | - |
alpha-D-glucose 1-phosphate | recombinant mutant E182A, pH 8.5, 27°C | Escherichia coli | |
0.2 | - |
UDP-glucose | recombinant wild-type, pH 8.5, 27°C | Escherichia coli | |
0.3 | - |
alpha-D-galactose 1-phosphate | recombinant wild-type, pH 8.5, 27°C | Escherichia coli | |
0.39 | - |
UDP-glucose | recombinant mutant E182A, pH 8.5, 27°C | Escherichia coli | |
0.77 | - |
alpha-D-galactose 1-phosphate | recombinant mutant E182A, pH 8.5, 27°C | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | 0.7 mol per mol of subunit | Escherichia coli | |
Fe2+ | not essential | Escherichia coli | |
Fe2+ | coordinated in a square pyramidal geometry with His296, His298, and Glu182 in a bidentate coordination providing the base ligands and His281 providing the axial ligand | Escherichia coli | |
additional information | metal content of mutants | Escherichia coli | |
Zn2+ | required | Escherichia coli | |
Zn2+ | 1.2 mol tightly bound per mol of subunit | Escherichia coli | |
Zn2+ | coordinated in a tetrahedral geometry by Cys52, Cys55, His115, and His164 | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P09148 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutants from strain BL21(DE3)pLysS to near homogeneity | Escherichia coli |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose | double-displacement mechanism | Escherichia coli | |
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose | active site triad: His164, His165, and His166, with His166 as nulceophilic catalyst, highly conserved | Escherichia coli |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.018 | - |
purified recombinant mutant H164N | Escherichia coli |
0.06 | - |
purified recombinant mutant C52S | Escherichia coli |
0.3 | - |
purified recombinant mutant C55S | Escherichia coli |
2.9 | - |
purified recombinant mutant H115N | Escherichia coli |
88.5 | - |
purified recombinant mutant E182A | Escherichia coli |
180 | - |
purified recombinant wild-type enzyme | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-glucose + alpha-D-galactose 1-phosphate | - |
Escherichia coli | alpha-D-glucose 1-phosphate + UDP-galactose | - |
r |
Synonyms | Comment | Organism |
---|---|---|
GALT | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
27 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0533 | - |
alpha-D-galactose 1-phosphate | recombinant mutant S161A, forward reaction, pH 8.5, 27°C | Escherichia coli | |
0.0967 | - |
UDP-galactose | recombinant mutant S161A, reverse reaction, pH 8.5, 27°C | Escherichia coli | |
55 | - |
UDP-galactose | recombinant mutant C160A, reverse reaction, pH 8.5, 27°C | Escherichia coli | |
177 | - |
UDP-galactose | recombinant mutant E182A, reverse reaction, pH 8.5, 27°C | Escherichia coli | |
265 | - |
alpha-D-galactose 1-phosphate | recombinant mutant C160A, forward reaction, pH 8.5, 27°C | Escherichia coli | |
283 | - |
UDP-galactose | recombinant wild-type, reverse reaction, pH 8.5, 27°C | Escherichia coli | |
653 | - |
alpha-D-galactose 1-phosphate | recombinant mutant E182A, forward reaction, pH 8.5, 27°C | Escherichia coli | |
780 | - |
alpha-D-galactose 1-phosphate | recombinant wild-type, forward reaction, pH 8.5, 27°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8.5 | - |
assay at | Escherichia coli |