Cloned (Comment) | Organism |
---|---|
expression of mutant H166G in strain CA13, expression of mutant H166A in strain BL21(DE3)pLysS | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
H166 A | site-directed mutagenesis, point mutation leads to shift of the enzyme activity to UDP-hexose synthase activity, formation of uridine 5'-phosphoimidazolate and alpha-D-glucose 1-phosphate from UDP-glucose and imidazole, highly reduced activity compared to H166G mutant | Escherichia coli |
H166G | site-directed mutagenesis | Escherichia coli |
H166G | point mutation leads to shift of the enzyme activity to UDP-hexose synthase activity, formation of uridine 5'-phosphoimidazolate and alpha-D-glucose 1-phosphate from UDP-glucose and imidazole | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | product inhibition study, wild-type and mutants H166G and H166A | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics | Escherichia coli | |
additional information | - |
additional information | mutant H166A | Escherichia coli | |
additional information | - |
additional information | mutant H166G | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | contains 0.67 mol per mol of wild-type enzyme, 0.59 mol per mol of mutant H166A enzyme, and 0.7-0.76 mol per mol of mutant H166G enzyme | Escherichia coli | |
Zn2+ | contains 1.21 mol per mol of wild-type enzyme, 1.33 mol per mol of mutant H166A enzyme, and 0.99-1.16 mol per mol of mutant H166G enzyme | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
native mutant H166G | - |
Purification (Comment) | Organism |
---|---|
recombinant mutants H166G and H166A | 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 | mutant H166G, random equilibrium, intrinsically ordered substrate binding mechanism, ping pong kinetics | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UDP-glucose + 2-methylimidazole | kinetic study | Escherichia coli | uridine 5'-phospho-2-methylimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + 2-methylimidazole | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | uridine 5'-phospho-2-methylimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + 4-methylimidazole | kinetic study | Escherichia coli | uridine 5'-phospho-4-methylimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + 4-methylimidazole | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | uridine 5'-phospho-4-methylimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + imidazole | mutant H166A | Escherichia coli | uridine 5'-phosphoimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + imidazole | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | uridine 5'-phosphoimidazole + alpha-D-glucose 1-phosphate | - |
r | |
UDP-glucose + imidazole | equilibrium study and constants | Escherichia coli | uridine 5'-phosphoimidazole + alpha-D-glucose 1-phosphate | - |
r | |
uridine 5'-phospho-2,4,5-trimethylimidazole + alpha-D-glucose 1-phosphate | kinetic study | Escherichia coli | UDP-glucose + 2,4,5-trimethylimidazole | - |
r | |
uridine 5'-phospho-2,4,5-trimethylimidazole + alpha-D-glucose 1-phosphate | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | UDP-glucose + 2,4,5-trimethylimidazole | - |
r | |
uridine 5'-phospho-2-methylimidazole + alpha-D-glucose 1-phosphate | kinetic study | Escherichia coli | UDP-glucose + 2-methylimidazole | - |
r | |
uridine 5'-phospho-2-methylimidazole + alpha-D-glucose 1-phosphate | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | UDP-glucose + 2-methylimidazole | - |
r | |
uridine 5'-phospho-3-methylimidazole + alpha-D-glucose 1-phosphate | kinetic study | Escherichia coli | UDP-glucose + 3-methylimidazole | - |
r | |
uridine 5'-phospho-3-methylimidazole + alpha-D-glucose 1-phosphate | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | UDP-glucose + 3-methylimidazole | - |
r | |
uridine 5'-phospho-4-methylimidazole + alpha-D-glucose 1-phosphate | kinetic study | Escherichia coli | UDP-glucose + 4-methylimidazole | - |
r | |
uridine 5'-phospho-4-methylimidazole + alpha-D-glucose 1-phosphate | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | UDP-glucose + 4-methylimidazole | - |
r | |
uridine 5'-phospho-5-methylimidazole + alpha-D-glucose 1-phosphate | kinetic study | Escherichia coli | UDP-glucose + 5-methylimidazole | - |
r | |
uridine 5'-phospho-5-methylimidazole + alpha-D-glucose 1-phosphate | mutant H166G, i.e. UDP-hexose synthase | Escherichia coli | UDP-glucose + 5-methylimidazole | - |
r |
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 |
---|---|---|---|---|---|
0.0167 | - |
imidazole | mutant H166A, forward reaction with UDP-glucose, pH 8.5, 27°C | Escherichia coli | |
0.0217 | - |
uridine 5'-phosphoimidazolate | mutant H166A, forward reaction with D-glucose 1-phosphate, pH 8.5, 27°C | Escherichia coli | |
5.52 | - |
imidazole | mutant H166G, forward reaction with UDP-glucose, pH 8.5, 27°C | Escherichia coli | |
13.5 | - |
uridine 5'-phosphoimidazolate | mutant H166G, forward reaction with D-glucose 1-phosphate, pH 8.5, 27°C | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
mutant H166G | Escherichia coli |
8.5 | - |
assay at | Escherichia coli |
8.5 | - |
both reaction directions | Escherichia coli |