Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutants in Escherichia coli JM109 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
C123S | site-directed mutagenesis, active enzyme, 7fold increase in Km for magnesium diphosphate, 2fold increased Ki for MgUTP2-, no longer sensitive to SH-reagents, e.g. iodoacetamide | Homo sapiens |
H266R | site-directed mutagenesis, mutant enzyme is active and similar to the wild-type, 4fold decrease in Km and Ki for MgUTP | Homo sapiens |
R389H | site-directed mutagenesis, mutant enzyme is active and similar to the wild-type | Homo sapiens |
R391K | site-directed mutagenesis, no activity, no correct folding | Homo sapiens |
R422Q | site-directed mutagenesis, mutant enzyme is active and similar to the wild-type | Homo sapiens |
R445H | site-directed mutagenesis, mutant enzyme is active and similar to the wild-type | Homo sapiens |
W218S | site-directed mutagenesis, mutant enzyme is active and similar to the wild-type, increase in Km | Homo sapiens |
W333S | site-directed mutagenesis, no activity, no correct folding | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
iodoacetamide | wild-type: loss of 56% activity after 30 min at 0.1 mM, mutant C123S is not affected | Homo sapiens | |
MgUTP | wild-type and mutants C123S, H266R, W218, R389H, R422Q, R445H | Homo sapiens | |
MgUTP2- | - |
Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.56 | - |
UTP | recombinant wild-type enzyme, pH 8.0, 25°C | Homo sapiens | |
0.56 | - |
UTP | UTP in form of MgUTP | Homo sapiens | |
0.68 | 1 | alpha-D-glucose 1-phosphate | recombinant wild-type enzyme, pH 8.0, 25°C | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type enzyme, partially from Escherichia coli JM109 | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
UTP + alpha-D-glucose 1-phosphate = diphosphate + UDP-glucose | Trp333 and Arg391 are essential for activity, while His266, Arg389, Arg422, Arg 445, and Trp218 are not | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
liver | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
diphosphate + CDP-glucose | very low activity, wild-type and mutant W218 | Homo sapiens | CTP + alpha-D-glucose 1-phosphate | - |
r | |
diphosphate + TDP-glucose | poor substrate | Homo sapiens | TTP + alpha-D-glucose 1-phosphate | - |
r | |
diphosphate + TDP-glucose | very low activity, wild-type and mutant W218 | Homo sapiens | TTP + alpha-D-glucose 1-phosphate | - |
r | |
UTP + alpha-D-glucose 1-phosphate | specific for alpha-D-glucose 1-phosphate | Homo sapiens | diphosphate + UDP-glucose | specific for UDP-glucose | r | |
UTP + alpha-D-glucose 1-phosphate | highly specific for UTP | Homo sapiens | diphosphate + UDP-glucose | specific for UDP-glucose | r |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.095 | - |
MgUTP2- | mutant H266R, pH 8.0, 25°C | Homo sapiens |