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Literature summary for 2.7.7.9 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.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
gene galU, expressio as His-tagged enzyme in Escherichia coli strain BL21(DE3) Helicobacter pylori

Crystallization (Commentary)

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

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

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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Helicobacter pylori O25363 gene galU
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Helicobacter pylori

Reaction

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 and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UTP + alpha-D-glucose 1-phosphate
-
Helicobacter pylori diphosphate + UDP-glucose
-
r

Subunits

Subunits Comment Organism
homotetramer the tetramerization of HpUGPase does not seem to be related to allosterism Helicobacter pylori

Synonyms

Synonyms Comment Organism
glucose-1-phosphate uridylyltransferase
-
Helicobacter pylori
UDP-glucose pyrophosphorylase
-
Helicobacter pylori
UGPase
-
Helicobacter pylori

General Information

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