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Literature summary for 2.7.7.12 extracted from

  • Geeganage, S.; Frey, P.A.
    Significance of metal ions in galactose-1-phosphate uridylyltransferase: an essential structural zinc and a nonessential structural iron (1999), Biochemistry, 38, 13398-13406.
    View publication on PubMed
Search for more literature for 2.7.7.12

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
C52S site-directed mutagenesis, 3000fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level Escherichia coli
C55S site-directed mutagenesis, 600fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level Escherichia coli
E182A site-directed mutagenesis, 50% activity compared to wild-type, normal formation of reaction intermediate UMP-enzyme, contains reduced zinc content and no iron Escherichia coli
H115N site-directed mutagenesis, 2.9% activity compared to wild-type, slightly reduced formation of reaction intermediate UMP-enzyme, retention of zinc and iron Escherichia coli
H164N site-directed mutagenesis, 10000fold less active than the wild-type, no formation of reaction intermediate UMP-enzyme, low metal content, low expression level Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
1,10-phenanthroline mutant E182A Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information
-
 Escherichia coli
0.048
-
 UDP-galactose recombinant mutant E182A, pH 8.5, 27°C Escherichia coli
0.12
-
 UDP-galactose recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.16
-
 alpha-D-glucose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.19
-
 alpha-D-glucose 1-phosphate recombinant mutant E182A, pH 8.5, 27°C Escherichia coli
0.2
-
 UDP-glucose recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.3
-
 alpha-D-galactose 1-phosphate recombinant wild-type, pH 8.5, 27°C Escherichia coli
0.39
-
 UDP-glucose recombinant mutant E182A, pH 8.5, 27°C Escherichia coli
0.77
-
 alpha-D-galactose 1-phosphate recombinant mutant E182A, pH 8.5, 27°C Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
Fe2+ 0.7 mol per mol of subunit Escherichia coli
Fe2+ not essential Escherichia coli
Fe2+ coordinated in a square pyramidal geometry with His296, His298, and Glu182 in a bidentate coordination providing the base ligands and His281 providing the axial ligand Escherichia coli
additional information metal content of mutants Escherichia coli
Zn2+ required Escherichia coli
Zn2+ 1.2 mol tightly bound per mol of subunit Escherichia coli
Zn2+ coordinated in a tetrahedral geometry by Cys52, Cys55, His115, and His164 Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli P09148
-
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose double-displacement mechanism Escherichia coli
a
UDP-alpha-D-glucose + alpha-D-galactose 1-phosphate = alpha-D-glucose 1-phosphate + UDP-alpha-D-galactose active site triad: His164, His165, and His166, with His166 as nulceophilic catalyst, highly conserved Escherichia coli
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.018
-
 purified recombinant mutant H164N Escherichia coli
0.06
-
 purified recombinant mutant C52S Escherichia coli
0.3
-
 purified recombinant mutant C55S Escherichia coli
2.9
-
 purified recombinant mutant H115N Escherichia coli
88.5
-
 purified recombinant mutant E182A Escherichia coli
180
-
 purified recombinant wild-type enzyme Escherichia coli

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

Synonyms

Synonyms Comment Organism
GALT
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
27
-
 assay at Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0533
-
 alpha-D-galactose 1-phosphate recombinant mutant S161A, forward reaction, pH 8.5, 27°C Escherichia coli
0.0967
-
 UDP-galactose recombinant mutant S161A, reverse reaction, pH 8.5, 27°C Escherichia coli
55
-
 UDP-galactose recombinant mutant C160A, reverse reaction, pH 8.5, 27°C Escherichia coli
177
-
 UDP-galactose recombinant mutant E182A, reverse reaction, pH 8.5, 27°C Escherichia coli
265
-
 alpha-D-galactose 1-phosphate recombinant mutant C160A, forward reaction, pH 8.5, 27°C Escherichia coli
283
-
 UDP-galactose recombinant wild-type, reverse reaction, pH 8.5, 27°C Escherichia coli
653
-
 alpha-D-galactose 1-phosphate recombinant mutant E182A, forward reaction, pH 8.5, 27°C Escherichia coli
780
-
 alpha-D-galactose 1-phosphate recombinant wild-type, forward reaction, pH 8.5, 27°C Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.5
-
 assay at Escherichia coli