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Literature summary for 2.7.7.12 extracted from

  • Quimby, B.B.; Wells, L.; Wilkinson, K.D.; Fridovich-Keil, J.L.
    Functional requirements of the active site position 185 in the human enzyme galactose-1-phosphate uridylyltransferase (1996), J. Biol. Chem., 271, 26835-26842.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutants in yeast Homo sapiens

Protein Variants

Protein Variants Comment Organism
P185A site-directed mutagenesis, reduced activity and reduced expression level compared to wild-type Homo sapiens
P185C site-directed mutagenesis, no remaining activity, same expression level compared to wild-type Homo sapiens
P185D site-directed mutagenesis, no remaining activity, same expression level compared to wild-type Homo sapiens
P185E site-directed mutagenesis, reduced activity, same expression level compared to wild-type Homo sapiens
P185F site-directed mutagenesis, no remaining activity, highly reduced expression level compared to wild-type Homo sapiens
P185G site-directed mutagenesis, reduced activity and expression level compared to wild-type Homo sapiens
P185H site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens
P185I site-directed mutagenesis, no remaining activity, highly reduced expression level compared to wild-type Homo sapiens
P185K site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens
P185L site-directed mutagenesis, no remaining activity, highly reduced expression level compared to wild-type Homo sapiens
P185M site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens
P185N site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens
P185Q site-directed mutagenesis, reduced activity, increased expression level compared to wild-type Homo sapiens
P185R site-directed mutagenesis, no remaining activity, same expression level compared to wild-type Homo sapiens
P185S site-directed mutagenesis, reduced activity, reduced expression level compared to wild-type Homo sapiens
P185T site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens
P185V site-directed mutagenesis, no remaining activity, highly reduced expression level compared to wild-type Homo sapiens
P185W site-directed mutagenesis, no remaining activity, highly reduced expression level compared to wild-type Homo sapiens
P185Y site-directed mutagenesis, no remaining activity, reduced expression level compared to wild-type Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
additional information kinetics Escherichia coli
additional information
-
additional information kinetics Homo sapiens
additional information
-
additional information wild-type and mutants in the 2-step reaction Escherichia coli
additional information
-
additional information wild-type and mutants in the 2-step reaction Homo sapiens
0.09
-
alpha-D-galactose 1-phosphate recombinant mutants P185E, pH 8.7, 37°C Homo sapiens
0.14
-
UDP-glucose recombinant mutant P185S, pH 8.7, 37°C Homo sapiens
0.15
-
alpha-D-galactose 1-phosphate recombinant mutants P185Q, pH 8.7, 37°C Homo sapiens
0.2
-
alpha-D-galactose 1-phosphate recombinant mutants P185G, pH 8.7, 37°C Homo sapiens
0.2
-
UDP-glucose recombinant wild-type and mutants P185E and P185A, pH 8.7, 37°C Homo sapiens
0.31
-
alpha-D-galactose 1-phosphate recombinant mutants P185S, pH 8.7, 37°C Homo sapiens
0.34
-
UDP-glucose recombinant mutants P185G, pH 8.7, 37°C Homo sapiens
0.4
-
UDP-glucose recombinant mutants P185Q, pH 8.7, 37°C Homo sapiens
0.42
-
alpha-D-galactose 1-phosphate recombinant mutants P185A, pH 8.7, 37°C Homo sapiens
0.57
-
alpha-D-galactose 1-phosphate recombinant wild-type, pH 8.7, 37°C Homo sapiens

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Homo sapiens
-
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged wild-type and mutants P185A, P185S, P185G, P185E, and P185Q to near homogeneity Homo sapiens

Reaction

Reaction Comment Organism Reaction ID
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose Pro185 has a critical role in facilitating the transferase reaction Escherichia coli
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose Pro185 has a critical role in facilitating the transferase reaction Homo sapiens

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.6
-
purified recombinant mutant P185E Homo sapiens
2.6
-
purified recombinant mutant P185Q Homo sapiens
6.14
-
purified recombinant mutant P185S Homo sapiens
6.9
-
purified recombinant mutant P185G Homo sapiens
9.3
-
purified recombinant mutant P185A Homo sapiens
122
-
purified recombinant wild-type enzyme Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
UDP-glucose + alpha-D-galactose 1-phosphate
-
Escherichia coli alpha-D-glucose 1-phosphate + UDP-galactose
-
r
UDP-glucose + alpha-D-galactose 1-phosphate
-
Homo sapiens alpha-D-glucose 1-phosphate + UDP-galactose
-
r

Synonyms

Synonyms Comment Organism
GALT
-
Homo sapiens

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
33
-
recombinant mutants P185G and P185A Homo sapiens
35
-
recombinant mutant P185S Homo sapiens
37
-
assay at Homo sapiens
40
-
recombinant mutants P185E, and P185Q Homo sapiens
45
-
recombinant wild-type Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.7
-
assay at Homo sapiens