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Literature summary 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)

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

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.7.7.12 additional information
-
 additional information kinetics Escherichia coli
2.7.7.12 additional information
-
 additional information kinetics Homo sapiens
2.7.7.12 additional information
-
 additional information wild-type and mutants in the 2-step reaction Escherichia coli
2.7.7.12 additional information
-
 additional information wild-type and mutants in the 2-step reaction Homo sapiens
2.7.7.12 0.09
-
 alpha-D-galactose 1-phosphate recombinant mutants P185E, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.14
-
 UDP-glucose recombinant mutant P185S, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.15
-
 alpha-D-galactose 1-phosphate recombinant mutants P185Q, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.2
-
 alpha-D-galactose 1-phosphate recombinant mutants P185G, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.2
-
 UDP-glucose recombinant wild-type and mutants P185E and P185A, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.31
-
 alpha-D-galactose 1-phosphate recombinant mutants P185S, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.34
-
 UDP-glucose recombinant mutants P185G, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.4
-
 UDP-glucose recombinant mutants P185Q, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.42
-
 alpha-D-galactose 1-phosphate recombinant mutants P185A, pH 8.7, 37°C Homo sapiens
2.7.7.12 0.57
-
 alpha-D-galactose 1-phosphate recombinant wild-type, pH 8.7, 37°C Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.12 Escherichia coli
-
-
-
2.7.7.12 Homo sapiens
-
-
-

Purification (Commentary)

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

Reaction

EC Number Reaction Comment Organism Reaction ID
2.7.7.12 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
a
2.7.7.12 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
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.7.7.12 1.6
-
 purified recombinant mutant P185E Homo sapiens
2.7.7.12 2.6
-
 purified recombinant mutant P185Q Homo sapiens
2.7.7.12 6.14
-
 purified recombinant mutant P185S Homo sapiens
2.7.7.12 6.9
-
 purified recombinant mutant P185G Homo sapiens
2.7.7.12 9.3
-
 purified recombinant mutant P185A Homo sapiens
2.7.7.12 122
-
 purified recombinant wild-type enzyme Homo sapiens

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
2.7.7.12 GALT
-
 Homo sapiens

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.12 33
-
 recombinant mutants P185G and P185A Homo sapiens
2.7.7.12 35
-
 recombinant mutant P185S Homo sapiens
2.7.7.12 37
-
 assay at Homo sapiens
2.7.7.12 40
-
 recombinant mutants P185E, and P185Q Homo sapiens
2.7.7.12 45
-
 recombinant wild-type Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.12 8.7
-
 assay at Homo sapiens