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Literature summary extracted from

  • Geeganage, S.; Ling, V.W.; Frey, P.A.
    Roles of two conserved amino acid residues in the active site of galactose-1-phosphate uridylyltransferase: an essential serine and a nonessential cysteine (2000), Biochemistry, 39, 5397-5404.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.12 overexpression of wild-type and mutants in strain BL21(DE3)pLysS Escherichia coli

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.12 C160A site-directed mutagenesis, 18fold reduced activity Escherichia coli
2.7.7.12 C160S site-directed mutagenesis, 12fold reduced activity Escherichia coli
2.7.7.12 H166G active site structure, complex of enzyme with UDP-D-galactose Escherichia coli
2.7.7.12 S161A site-directed mutagenesis, 7000fold reduced activity compared to wild-type Escherichia coli

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 at 4°C, wild-type and mutants Escherichia coli
2.7.7.12 0.062
-
 UDP-galactose recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.072
-
 UDP-glucose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.078
-
 UDP-galactose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.082
-
 UDP-glucose recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.12
-
 UDP-galactose recombinant wild-type, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.125
-
 alpha-D-galactose 1-phosphate recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.14
-
 alpha-D-glucose 1-phosphate recombinant mutant C160A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.16
-
 alpha-D-glucose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.2
-
 alpha-D-glucose 1-phosphate recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.2
-
 UDP-glucose recombinant wild-type, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.223
-
 alpha-D-galactose 1-phosphate recombinant mutant S161A, pH 8.5, 27°C Escherichia coli
2.7.7.12 0.3
-
 alpha-D-galactose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
2.7.7.12 Fe2+ 0.7 mol per mol of subunit Escherichia coli
2.7.7.12 Fe2+ mutant C160A: 0.5 mol per mol of subunit, mutant S161A: 0.4 mol per mol of subunit Escherichia coli
2.7.7.12 Zn2+ 1.2 mol tightly bound per mol of subunit Escherichia coli
2.7.7.12 Zn2+ mutant C160A: 1.3 mol per mol of subunit, mutant S161A: 1.2 mol per mol of subunit like the wild-type Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.12 Escherichia coli P09148 galT gene
-

Purification (Commentary)

EC Number Purification (Comment) Organism
2.7.7.12 recombinant wild-type and mutants from strain BL21(DE3)pLysS to near homogeneity Escherichia coli

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 double-displacement mechanism 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 active site structure, enzyme complexed with UDP-galactose Escherichia coli
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 0.00093
-
 recombinant mutant S161A, cell extract Escherichia coli
2.7.7.12 0.023
-
 purified recombinant mutant S161A Escherichia coli
2.7.7.12 9.9
-
 recombinant mutant C160A, cell extract Escherichia coli
2.7.7.12 14.9
-
 recombinant mutant C160S, cell extract Escherichia coli
2.7.7.12 16.3
-
 recombinant wild-type, cell extract Escherichia coli
2.7.7.12 79.5
-
 purified recombinant mutant C160A Escherichia coli
2.7.7.12 180
-
 purified recombinant wild-type enzyme Escherichia coli

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

Synonyms

EC Number Synonyms Comment Organism
2.7.7.12 GALT
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.7.7.12 283
-
 UDP-galactose recombinant wild-type, reverse reaction, pH 8.5, 27°C Escherichia coli
2.7.7.12 780
-
 alpha-D-galactose 1-phosphate recombinant wild-type, forward reaction, pH 8.5, 27°C Escherichia coli