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Literature summary for 2.7.1.6 extracted from

  • McAuley, M.; Huang, M.; Timson, D.J.
    Insight into the mechanism of galactokinase Role of a critical glutamate residue and helix/coil transitions (2017), Biochim. Biophys. Acta Proteins Proteom., 1865, 321-328 .
    View publication on PubMed
Search for more literature for 2.7.1.6

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli HMS174(DE3) Homo sapiens

Protein Variants

Protein Variants Comment Organism
E174Q no detectable activity, less stable enzyme Homo sapiens
R105M modest reduction in activity with little change in stability Homo sapiens
R228M interaction with both ATP and galactose is affected. The variant is significantly less stable than the wild-type protein Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0039
-
 ATP mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
0.004
-
 ATP wild-type enzyme, pH 7.5, 37°C Homo sapiens
0.0094
-
 ATP mutant enzyme R228M, pH 7.5, 37°C Homo sapiens
0.0149
-
 ATP mutant enzyme R228K, pH 7.5, 37°C Homo sapiens
0.083
-
 ATP mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
0.22
-
 alpha-D-galactose mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
0.26
-
 alpha-D-galactose mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
0.31
-
 alpha-D-galactose mutant enzyme R228K, pH 7.5, 37°C Homo sapiens
0.33
-
 alpha-D-galactose wild-type enzyme, pH 7.5, 37°C Homo sapiens
0.67
-
 alpha-D-galactose mutant enzyme R228M, pH 7.5, 37°C Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + alpha-D-galactose Homo sapiens the enzyme catalyses the first committed step in the Leloir pathway ADP + alpha-D-galactose 1-phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens P51570
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + alpha-D-galactose the enzyme catalyses the first committed step in the Leloir pathway Homo sapiens ADP + alpha-D-galactose 1-phosphate
-
 ?
ATP + alpha-D-galactose negative charge at residue 174 is essential for catalysis Homo sapiens ADP + alpha-D-galactose 1-phosphate
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
51.3
-
 Tm-value, mutant enzyme R228K Homo sapiens
51.68
-
 Tm-value, mutant enzyme R228M Homo sapiens
52.5
-
 Tm-value, mutant enzyme E174Q Homo sapiens
55.13
-
 Tm-value, mutant enzyme E174D Homo sapiens
55.42
-
 Tm-value, mutant enzyme R105M Homo sapiens
55.72
-
 Tm-value, wild-type enzyme Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.7
-
 ATP mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
0.7
-
 alpha-D-galactose mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
1.6
-
 ATP mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
1.6
-
 alpha-D-galactose mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
1.7
-
 ATP wild-type enzyme, pH 7.5, 37°C Homo sapiens
1.7
-
 alpha-D-galactose wild-type enzyme, pH 7.5, 37°C Homo sapiens
2.3
-
 ATP mutant enzyme R228M, pH 7.5, 37°C Homo sapiens
2.3
-
 alpha-D-galactose mutant enzyme R228M, pH 7.5, 37°C Homo sapiens
3.7
-
 ATP mutant enzyme R228K, pH 7.5, 37°C Homo sapiens
3.7
-
 alpha-D-galactose mutant enzyme R228K, pH 7.5, 37°C Homo sapiens

General Information

General Information Comment Organism
drug target possible drug target in the treatment of type I galactosemia Homo sapiens
metabolism the enzyme catalyses the first committed step in the Leloir pathway Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
2.7
-
 alpha-D-galactose mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
3.3
-
 alpha-D-galactose mutant enzyme R228M, pH 7.5, 37°C Homo sapiens
7.6
-
 alpha-D-galactose mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
10.1
-
 alpha-D-galactose wild-type enzyme, pH 7.5, 37°C Homo sapiens
19.6
-
 alpha-D-galactose mutant enzyme R228K, pH 7.5, 37°C Homo sapiens
98
-
 ATP mutant enzyme R105M, pH 7.5, 37°C Homo sapiens
240
-
 ATP mutant enzyme R228M, pH 7.5, 37°C Homo sapiens
250
-
 ATP mutant enzyme R228K, pH 7.5, 37°C Homo sapiens
340
-
 ATP mutant enzyme E174D, pH 7.5, 37°C Homo sapiens
430
-
 ATP wild-type enzyme, pH 7.5, 37°C Homo sapiens