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Literature summary extracted from

  • Hyde, A.S.; Thelen, A.M.; Barycki, J.J.; Simpson, M.A.
    UDP-glucose dehydrogenase activity and optimal downstream cellular function require dynamic reorganization at the dimer-dimer subunit interfaces (2013), J. Biol. Chem., 288, 35049-35057.
    View publication on PubMedView publication on EuropePMC

Application

EC Number Application Comment Organism
1.1.1.22 medicine manipulation of UGDH activity by hexamer stabilization may offer therapeutic options in cancer and other pathologies Homo sapiens

Cloned(Commentary)

EC Number Cloned (Comment) Organism
1.1.1.22 recombinant expression of N-terminally His6-tagged wild-type and mutant enzymes in Escherichia coli strain Rosetta2(DE3)pLysS, recombinant expression in HEK293 cells Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.1.1.22 E110A site-directed mutagenesis, the mutant, although dimeric in the apo form, exhibits only about 50% reduction in Vmax, but is highly unstable in solution and in cultured cells so it cannot be evaluated unambiguously Homo sapiens
1.1.1.22 additional information UDP-glucose dehydrogenase mutants are engineered to perturb hexamer:dimer quaternary structure equilibrium. Dimeric species of UGDH have reduced activity in vitro and in supporting hyaluronan production by cultured cells. The purified enzymes reveal a significant decrease in the enzymatic activity of the obligate dimer and hexamer mutants. The activity of the truncated DELTA132 mutant is negligible. The half-life of UGDH catalytic activity in vitro is reduced by mutations at the dimer interface Homo sapiens
1.1.1.22 T325A site-directed mutagenesis, the mutant occurs as dimeric species that can be induced to form hexamers in the ternary complex with substrate and cofactor. The inducible hexamer shows that upon increasing enzyme concentration, which favors the hexameric species, activity is modestly decreased and exhibits cooperativity. The T325A mutant is significantly less efficient in promoting downstream hyaluronan production by HEK293 cells than the wild-type. The activity of the T325A mutant is the most labile, with a half-life of only 24 h that is not extended significantly by substrate and cofactor addition Homo sapiens
1.1.1.22 T325D site-directed mutagenesis, the mutant yields exclusively dimeric species. The T325D mutant is significantly less efficient in promoting downstream hyaluronan production by HEK293 cells than the wild-type. UGDH T325D retains its activity similarly to the wild-type enzyme but does not exhibit increased stability in the abortive ternary complex Homo sapiens

General Stability

EC Number General Stability Organism
1.1.1.22 the half-life of UGDH catalytic activity in vitro is reduced by mutations at the dimer interface Homo sapiens

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.1.1.22 UDP-xylose a potent UGDH inhibitor Homo sapiens

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.1.1.22 additional information
-
 additional information Michaelis-Menten kinetic, cooperative kinetic behavior occurs in the hexameric enzyme Homo sapiens
1.1.1.22 0.015
-
 UDP-alpha-D-glucose pH 7.4, 37°C, recombinant His-tagged mutant T325D Homo sapiens
1.1.1.22 0.034
-
 UDP-alpha-D-glucose pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Homo sapiens
1.1.1.22 0.048
-
 UDP-alpha-D-glucose pH 7.4, 37°C, recombinant His-tagged mutant T325A Homo sapiens
1.1.1.22 0.646
-
 NAD+ pH 7.4, 37°C, recombinant His-tagged wild-type enzyme Homo sapiens
1.1.1.22 0.897
-
 NAD+ pH 7.4, 37°C, recombinant His-tagged mutant T325A Homo sapiens
1.1.1.22 1.084
-
 NAD+ pH 7.4, 37°C, recombinant His-tagged mutant T325D Homo sapiens

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.1.1.22 57000
-
 x * 57000, recombinant enzyme, SDS-PAGE Homo sapiens

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
1.1.1.22 additional information Homo sapiens the transient capacity to dissociate and reorganize the hydrogen bond network at the interface between dimeric units is an important element of the normal catalytic cycle ?
-
 ?
1.1.1.22 UDP-alpha-D-glucose + 2 NAD+ + H2O Homo sapiens
-
 UDP-alpha-D-glucuronate + 2 NADH + 2 H+
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
1.1.1.22 Homo sapiens O60701
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.1.1.22 recombinant N-terminally His6-tagged wild-type and mutant enzymes from Escherichia coli strain Rosetta2(DE3)pLysS by nickel affinity chromatography and dialysis Homo sapiens

Source Tissue

EC Number Source Tissue Comment Organism Textmining
1.1.1.22 cell culture
-
 Homo sapiens
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.1.1.22 additional information the transient capacity to dissociate and reorganize the hydrogen bond network at the interface between dimeric units is an important element of the normal catalytic cycle Homo sapiens ?
-
 ?
1.1.1.22 UDP-alpha-D-glucose + 2 NAD+ + H2O
-
 Homo sapiens UDP-alpha-D-glucuronate + 2 NADH + 2 H+
-
 ?

Subunits

EC Number Subunits Comment Organism
1.1.1.22 dimer or hexamer x * 57000, recombinant enzyme, SDS-PAGE Homo sapiens
1.1.1.22 More examination of the dimer-dimer subunit interface reveals an extensive network of charge interactions and hydrogen bonding that coordinately stabilize the hexamer in the presence and absence of its cofactor or substrate, involving residue T325. The wild-type UGDH enzyme purifies in a hexamer-dimer equilibrium and transiently undergoes dynamic motion that exposes the dimer-dimer interface during catalysis. Only dynamic UGDH hexamers support robust cellular function, mutant dimeric species of UGDH have reduced activity in vitro and in supporting hyaluronan production by cultured cells. Molecular interactions at the subunit interface, overview. In the apo form Thr325 directly forms a hydrogen bond with Asp105 of the opposite subunit Homo sapiens

Synonyms

EC Number Synonyms Comment Organism
1.1.1.22 UDP-glucose dehydrogenase
-
 Homo sapiens
1.1.1.22 UGDH
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.1.1.22 37
-
 assay at Homo sapiens

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
1.1.1.22 additional information
-
 thermal stabilities of wild-type and mutant enzymes, overview Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.1.1.22 7.4
-
 assay at Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
1.1.1.22 NAD+
-
 Homo sapiens

General Information

EC Number General Information Comment Organism
1.1.1.22 malfunction mutant dimeric species of UGDH have reduced activity in vitro and in supporting hyaluronan production by cultured cells. The purified enzymes reveal a significant decrease in the enzymatic activity of the obligate dimer and hexamer mutants. Both T325A and T325D mutants were significantly less efficient in promoting downstream hyaluronan production by HEK293 cells Homo sapiens
1.1.1.22 physiological function UDP-glucose dehydrogenase activity and optimal downstream cellular function require dynamic reorganization at the dimer-dimer subunit interfaces Homo sapiens