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Literature summary for 2.7.7.9 extracted from

  • Chang, H.Y.; Peng, H.L.; Chao, Y.C.; Duggleby, R.G.
    The importance of conserved residues in human liver UDPglucose pyrophosphorylase (1996), Eur. J. Biochem., 236, 723-728.
    View publication on PubMed
Search for more literature for 2.7.7.9

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutants in Escherichia coli JM109 Homo sapiens

Protein Variants

Protein Variants Comment Organism
C123S site-directed mutagenesis, active enzyme, 7fold increase in Km for magnesium diphosphate, 2fold increased Ki for MgUTP2-, no longer sensitive to SH-reagents, e.g. iodoacetamide Homo sapiens
H266R site-directed mutagenesis, mutant enzyme is active and similar to the wild-type, 4fold decrease in Km and Ki for MgUTP Homo sapiens
R389H site-directed mutagenesis, mutant enzyme is active and similar to the wild-type Homo sapiens
R391K site-directed mutagenesis, no activity, no correct folding Homo sapiens
R422Q site-directed mutagenesis, mutant enzyme is active and similar to the wild-type Homo sapiens
R445H site-directed mutagenesis, mutant enzyme is active and similar to the wild-type Homo sapiens
W218S site-directed mutagenesis, mutant enzyme is active and similar to the wild-type, increase in Km Homo sapiens
W333S site-directed mutagenesis, no activity, no correct folding Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
iodoacetamide wild-type: loss of 56% activity after 30 min at 0.1 mM, mutant C123S is not affected Homo sapiens
MgUTP wild-type and mutants C123S, H266R, W218, R389H, R422Q, R445H Homo sapiens
MgUTP2-
-
 Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.56
-
 UTP recombinant wild-type enzyme, pH 8.0, 25°C Homo sapiens
0.56
-
 UTP UTP in form of MgUTP Homo sapiens
0.68 1 alpha-D-glucose 1-phosphate recombinant wild-type enzyme, pH 8.0, 25°C Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type enzyme, partially from Escherichia coli JM109 Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
UTP + alpha-D-glucose 1-phosphate = diphosphate + UDP-glucose Trp333 and Arg391 are essential for activity, while His266, Arg389, Arg422, Arg 445, and Trp218 are not Homo sapiens
a

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
 Homo sapiens
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
diphosphate + CDP-glucose very low activity, wild-type and mutant W218 Homo sapiens CTP + alpha-D-glucose 1-phosphate
-
 r
diphosphate + TDP-glucose poor substrate Homo sapiens TTP + alpha-D-glucose 1-phosphate
-
 r
diphosphate + TDP-glucose very low activity, wild-type and mutant W218 Homo sapiens TTP + alpha-D-glucose 1-phosphate
-
 r
UTP + alpha-D-glucose 1-phosphate specific for alpha-D-glucose 1-phosphate Homo sapiens diphosphate + UDP-glucose specific for UDP-glucose r
UTP + alpha-D-glucose 1-phosphate highly specific for UTP Homo sapiens diphosphate + UDP-glucose specific for UDP-glucose r

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.095
-
 MgUTP2- mutant H266R, pH 8.0, 25°C Homo sapiens