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Literature summary for 2.7.7.9 extracted from

  • Yu, Q.; Zheng, X.
    The crystal structure of human UDP-glucose pyrophosphorylase reveals a latch effect that influences enzymatic activity (2012), Biochem. J., 442, 283-291.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Saccharomyces cerevisiae
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Drosophila melanogaster
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Danio rerio
expression of His-tagged enzyme in Escherichia coli strain BL21(DE3) Caenorhabditis elegans
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Homo sapiens

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme, hanging drop vapour diffusion method, mixing of 0.002 ml of 10 mg/ml protein in 20 mM Tris/HCl, pH 8.0, and 200 mM NaCl, with 0.001 ml of reservoir solution containing 100 mM HEPES, pH 6.5, 5 mM MgSO4, 15% w/v PEG 3350 and 20% v/v glycerol, 20°C, X-ray diffraction structure determination and analysis at 3.6 A resolution, molecular replacement Homo sapiens

Protein Variants

Protein Variants Comment Organism
E412D site-directed mutagenesis, the mutation does not change the oligomeric state, the mutant shows 176% catalytic activity compared to the wild-type enzyme Homo sapiens
E412K site-directed mutagenesis, the mutant has a longer side chain with a reverse in charge showed obvious inhibitory effects which results in 78% reduced activity compared to the wild-type hUGPase activity Homo sapiens
E412Q site-directed mutagenesis, the mutation changes the charge property, but not the length of side chain and shows only a marginal increase in activity of 19% when compared with the wild-type protein Homo sapiens
K4110S site-directed mutagenesis, the mutant shows increased activity compared to the wild-type enzyme Homo sapiens
N491P/L492E site-directed mutagenesis, mutant N491P/L492E is constructed to depolymerize hUGPase octamers, the mutation in the C-terminal left-handed beta-helix changes the oligomerization state the mutant enzyme, that becomes monomeric, it shows about the double activity of the wild-type enzyme Homo sapiens
P414G/T415P site-directed mutagenesis, the mutant shows activity similar to that of the wild-type hUGPase, the mutation does not change the oligomeric state Homo sapiens
S309N/S311R site-directed mutagenesis, mutation in sequence analogy to the Saccharomyces cerevisiae enzyme, the mutant shows 84% of wild-type activity Homo sapiens
T406K/M407L site-directed mutagenesis, the mutant shows activity similar to that of the wild-type hUGPase, the mutation does not change the oligomeric state Homo sapiens
V416N site-directed mutagenesis, the mutant shows activity similar to that of the wild-type hUGPase, the mutation does not change the oligomeric state Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.067
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant E412K Homo sapiens
0.151
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant K410S Homo sapiens
0.155
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant E412Q Homo sapiens
0.183
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant enzyme Danio rerio
0.198
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant N491P/L492E Homo sapiens
0.202
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant P414G/T415P Homo sapiens
0.205
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant E412D Homo sapiens
0.217
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant V416N Homo sapiens
0.219
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant T406K/M407L Homo sapiens
0.235
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant mutant S309N/S311R Homo sapiens
0.253
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant enzyme Homo sapiens
0.316
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant enzyme Drosophila melanogaster
0.715
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant enzyme Caenorhabditis elegans
0.83
-
alpha-D-glucose 1-phosphate pH 7.5, 37°C, recombinant enzyme Saccharomyces cerevisiae

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ required Homo sapiens
Mg2+ required Saccharomyces cerevisiae

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
55000
-
8 * 55000, SDS-PAGE, human UGPase forms octamers through end-to-end and side-by-side interactions, structure, overview Homo sapiens
55000
-
8 * 55000, SDS-PAGE, the C-terminal left-handed beta-helices are important for the formation of the yUGPase octamer Saccharomyces cerevisiae
79000
-
1 * 79000, mutant enzyme N491P/L492E, SDS-PAGE, depolymerization of hUGPase, like in mutant N491P/L492E, results in monomers and higher enzymatic activity Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
UTP + alpha-D-glucose 1-phosphate Homo sapiens
-
diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate Saccharomyces cerevisiae
-
diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate Drosophila melanogaster
-
diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate Danio rerio
-
diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate Caenorhabditis elegans
-
diphosphate + UDP-glucose
-
r

Organism

Organism UniProt Comment Textmining
Caenorhabditis elegans Q9XUS5
-
-
Danio rerio B8JMZ1
-
-
Drosophila melanogaster A5XCL5
-
-
Homo sapiens Q16851 isoform II
-
Saccharomyces cerevisiae C7GP37
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Saccharomyces cerevisiae
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Drosophila melanogaster
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Danio rerio
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Caenorhabditis elegans
recombinnat His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UTP + alpha-D-glucose 1-phosphate
-
Homo sapiens diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate
-
Saccharomyces cerevisiae diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate
-
Drosophila melanogaster diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate
-
Danio rerio diphosphate + UDP-glucose
-
r
UTP + alpha-D-glucose 1-phosphate
-
Caenorhabditis elegans diphosphate + UDP-glucose
-
r

Subunits

Subunits Comment Organism
homooctamer 8 * 55000, SDS-PAGE, human UGPase forms octamers through end-to-end and side-by-side interactions, structure, overview Homo sapiens
homooctamer 8 * 55000, SDS-PAGE, the C-terminal left-handed beta-helices are important for the formation of the yUGPase octamer Saccharomyces cerevisiae
monomer 1 * 79000, mutant enzyme N491P/L492E, SDS-PAGE, depolymerization of hUGPase, like in mutant N491P/L492E, results in monomers and higher enzymatic activity Homo sapiens
More structure comparison of human and yeast enzyme, overview Homo sapiens
More structure comparison of human and yeast enzyme, overview Saccharomyces cerevisiae
octamer
-
Drosophila melanogaster
octamer
-
Danio rerio
octamer
-
Caenorhabditis elegans

Synonyms

Synonyms Comment Organism
UDP-glucose pyrophosphorylase
-
Homo sapiens
UDP-glucose pyrophosphorylase
-
Saccharomyces cerevisiae
UDP-glucose pyrophosphorylase
-
Drosophila melanogaster
UDP-glucose pyrophosphorylase
-
Danio rerio
UDP-glucose pyrophosphorylase
-
Caenorhabditis elegans
UGPase
-
Homo sapiens
UGPase
-
Saccharomyces cerevisiae
UGPase
-
Drosophila melanogaster
UGPase
-
Danio rerio
UGPase
-
Caenorhabditis elegans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Homo sapiens
37
-
assay at Saccharomyces cerevisiae
37
-
assay at Drosophila melanogaster
37
-
assay at Danio rerio
37
-
assay at Caenorhabditis elegans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Homo sapiens
7.5
-
assay at Saccharomyces cerevisiae
7.5
-
assay at Drosophila melanogaster
7.5
-
assay at Danio rerio
7.5
-
assay at Caenorhabditis elegans

General Information

General Information Comment Organism
additional information structure comparison of human and yeast enzyme, overview Saccharomyces cerevisiae
additional information structure comparison of human and yeast enzyme, overview. Depolymerization of hUGPase, like in mutant N491P/L492E, results in monomers and higher enzymatic activity Homo sapiens
physiological function the enzyme is essential for survival Saccharomyces cerevisiae