Literature summary extracted from
Kim, H.; Choi, J.; Kim, T.; Lokanath, N.K.; Ha, S.C.; Suh, S.W.; Hwang, H.Y.; Kim, K.K.
Structural basis for the reaction mechanism of UDP-glucose pyrophosphorylase (2010), Mol. Cells, 29, 397-405.
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
2.7.7.9 |
gene galU, expressio as His-tagged enzyme in Escherichia coli strain BL21(DE3) |
Helicobacter pylori |
Crystallization (Commentary)
EC Number |
Crystallization (Comment) |
Organism |
---|
2.7.7.9 |
apo- and UDP-glucose/Mg2+-bound enzyme complexes, hanging drop and sitting drop vapor diffusion methods, protein in 20 mM Tris-HCl pH 7.5 and 0.1 M NaCl, is mixed with 0.1 M sodium acetate trihydrate, pH 4.6, 2 M ammonium sulfate and 0.1 M guanidine-HCl for the apo-enzyme crystals, 22°C, one week. UDP-Glc/Mg2+-bound holo-UGPase is crystallized in 0.1 M HEPES-Na, pH 7.5, 2% PEG 400 and 1.5 M ammonium sulfate containing 10 mM UDP-Glc and 10 mM MgCl2 at 22°C within a month, X-ray diffraction structure determination and analysis at 2.9 A and 2.3 A resolutions, respectively |
Helicobacter pylori |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
2.7.7.9 |
D130A |
site-directed mutagenesis the mutant shows highly reduced activity compatred to the wild-type enzyme |
Helicobacter pylori |
2.7.7.9 |
R15A |
site-directed mutagenesis the mutant shows highly reduced activity compatred to the wild-type enzyme |
Helicobacter pylori |
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
2.7.7.9 |
Mg2+ |
the Mg2+ ion coordinated by Asp130, two oxygen atoms of phosphoryl groups, and three water molecules with octahedral geometry. The Mg2+ ion plays a key role in the enzymatic activity of UGPase by enhancing the binding of UGPase to UTP or UDP-glucose |
Helicobacter pylori |
|
Natural Substrates/ Products (Substrates)
EC Number |
Natural Substrates |
Organism |
Comment (Nat. Sub.) |
Natural Products |
Comment (Nat. Pro.) |
Rev. |
Reac. |
---|
2.7.7.9 |
UTP + alpha-D-glucose 1-phosphate |
Helicobacter pylori |
- |
diphosphate + UDP-glucose |
- |
r |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
2.7.7.9 |
Helicobacter pylori |
O25363 |
gene galU |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
2.7.7.9 |
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration |
Helicobacter pylori |
Reaction
EC Number |
Reaction |
Comment |
Organism |
Reaction ID |
---|
2.7.7.9 |
UTP + alpha-D-glucose 1-phosphate = diphosphate + UDP-glucose |
ordered sequential Bi Bi mechanism, overview. Pyrimidine bases may approach the base binding site, but they cannot form the tight interactions observed in the HpUGPase/UDP-Glc complex. Thymine binding would be hindered by its extra methyl group, and cytosine could not make specific hydrogen bonds |
Helicobacter pylori |
|
Substrates and Products (Substrate)
EC Number |
Substrates |
Comment Substrates |
Organism |
Products |
Comment (Products) |
Rev. |
Reac. |
---|
2.7.7.9 |
UTP + alpha-D-glucose 1-phosphate |
- |
Helicobacter pylori |
diphosphate + UDP-glucose |
- |
r |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
2.7.7.9 |
homotetramer |
the tetramerization of HpUGPase does not seem to be related to allosterism |
Helicobacter pylori |
Synonyms
EC Number |
Synonyms |
Comment |
Organism |
---|
2.7.7.9 |
glucose-1-phosphate uridylyltransferase |
- |
Helicobacter pylori |
2.7.7.9 |
UDP-glucose pyrophosphorylase |
- |
Helicobacter pylori |
2.7.7.9 |
UGPase |
- |
Helicobacter pylori |
General Information
EC Number |
General Information |
Comment |
Organism |
---|
2.7.7.9 |
additional information |
structural basis for the reaction mechanism of UDP-glucose pyrophosphorylase, overview. The active sites are located in a deep pocket of each subunit. The tetramerization of HpUGPase does not seem to be related to allosterism |
Helicobacter pylori |
2.7.7.9 |
physiological function |
UGPase is crucial in carbohydrate metabolism since UDP-Glc is used for the biosynthesis of glycogen and many other carbohydrate derivatives such as glycolipids, glycoproteins and proteoglycans |
Helicobacter pylori |