Cloned (Comment) | Organism |
---|---|
gene galU, expressio as His-tagged enzyme in Escherichia coli strain BL21(DE3) | Helicobacter pylori |
Crystallization (Comment) | Organism |
---|---|
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 | Comment | Organism |
---|---|---|
D130A | site-directed mutagenesis the mutant shows highly reduced activity compatred to the wild-type enzyme | Helicobacter pylori |
R15A | site-directed mutagenesis the mutant shows highly reduced activity compatred to the wild-type enzyme | Helicobacter pylori |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
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 | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
UTP + alpha-D-glucose 1-phosphate | Helicobacter pylori | - |
diphosphate + UDP-glucose | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Helicobacter pylori | O25363 | gene galU | - |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration | Helicobacter pylori |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
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 | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
UTP + alpha-D-glucose 1-phosphate | - |
Helicobacter pylori | diphosphate + UDP-glucose | - |
r |
Subunits | Comment | Organism |
---|---|---|
homotetramer | the tetramerization of HpUGPase does not seem to be related to allosterism | Helicobacter pylori |
Synonyms | Comment | Organism |
---|---|---|
glucose-1-phosphate uridylyltransferase | - |
Helicobacter pylori |
UDP-glucose pyrophosphorylase | - |
Helicobacter pylori |
UGPase | - |
Helicobacter pylori |
General Information | Comment | Organism |
---|---|---|
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 |
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 |