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Literature summary for 2.7.1.6 extracted from

  • McAuley, M.; Huang, M.; Timson, D.J.
    Modulation of the mobility of a key region in human galactokinase Impacts on catalysis and stability (2018), Bioorg. Chem., 81, 649-657 .
    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
E244S significantly increased (5fold) Michaelis constant for ATP, indicating that this variation impacts on the interaction of the protein with ATP. Significantly increased Michaelis constants for galactose. Turnover number is significantly increased Homo sapiens
E245S catalyzes less efficiently the phosphorylation of galactose than the wild-type enzyme. Significantly increased Michaelis constants for galactose. Turnover number is significantly increased Homo sapiens
L231S catalyzes less efficiently the phosphorylation of galactose than the wild-type enzyme. Reduced turnover and decreased catalytic efficiency with the substrates 2-deoxy-D-galactose and ATP Homo sapiens
Q242S the turnover number and the catalytic efficiency with ATP (cosubstrate 2-deoxy-D-galactose) are increased. Inactive with D-galactosamine. Turnover number is significantly increased Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0024
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
0.0039
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.0066
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.0077
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.0094
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
0.0128
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
0.0132
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.0148
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
0.0162
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.0198
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.0367
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.0392
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.0432
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.0446
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
0.0594
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.0594
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.162
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.17
-
 alpha-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.26
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
0.44
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
0.71
-
 D-talose pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.71
-
 D-galactosamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
0.83
-
 D-talose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.92
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.92
-
 D-talose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.97
-
 2-deoxy-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
1
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
1.1
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
1.1
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
1.8
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
1.9
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
2.3
-
 D-talose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
2.7
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
2.9
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E245S 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
-
 ADP + alpha-D-galactose 1-phosphate
-
 ?

Organic Solvent Stability

Organic Solvent Comment Organism
urea the wild-type protein gives a discrete band up to 1 M urea. Mutant enzyme L231S behaves in a similar manner. Q242S is stable up to 0.5 M urea. E244S and E245S are unstable under these conditions even in the absence of urea, indicating a large effect of these variants on the protein, consistent with the molecular dynamics analysis of these two variants Homo sapiens

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 + 2-deoxy-D-galactose
-
 Homo sapiens ADP + 2-deoxy-alpha-D-galactose 1-phosphate
-
 ?
ATP + alpha-D-galactose
-
 Homo sapiens ADP + alpha-D-galactose 1-phosphate
-
 ?
ATP + D-galactosamine
-
 Homo sapiens ADP + D-galactosamine 1-phosphate
-
 ?
ATP + D-talose
-
 Homo sapiens ADP + alpha-D-talose 1-phosphate
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
52.7
-
 Tm-value, mutant enzyme Q242S Homo sapiens
54
-
 Tm-value, mutant enzyme E245S Homo sapiens
55.1
-
 Tm-value, mutant enzyme E244S Homo sapiens
55.8
-
 Tm-value, mutant enzyme L231S Homo sapiens
56.1
-
 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
0.4
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
0.4
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
0.5
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.5
-
 D-talose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
1
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
1
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
1.7
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
1.7
-
 alpha-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
1.7
-
 D-talose pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.1
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.1
-
 D-galactosamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
3.4
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
3.4
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
3.4
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
3.4
-
 D-talose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
4
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
4
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
5.9
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
5.9
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
6.1
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
6.1
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
6.8
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
6.8
-
 2-deoxy-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
8
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
8
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
11
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
11
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
15
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
15
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
18.1
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
18.1
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
18.1
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
18.1
-
 D-talose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.4
-
 D-talose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
0.4
-
 D-talose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
0.5
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
1.2
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
1.5
-
 D-talose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
2.9
-
 D-talose pH 7.5, 37°C, wild-type enzyme Homo sapiens
2.9
-
 D-galactosamine pH 7.5, 37°C, wild-type enzyme Homo sapiens
3.9
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
3.9
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
4.5
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
5.6
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
5.8
-
 2-deoxy-D-galactose pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
6.2
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
7.1
-
 2-deoxy-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
10.1
-
 alpha-D-galactose pH 7.5, 37°C, wild-type enzyme Homo sapiens
13.5
-
 alpha-D-galactose pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
19.7
-
 D-galactosamine pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
20
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
30
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
73
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
110
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
190
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
210
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
300
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
310
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
320
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
360
-
 ATP pH 7.5, 37°C, mutant enzyme E244S Homo sapiens
360
-
 ATP pH 7.5, 37°C, mutant enzyme L231S Homo sapiens
380
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
420
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens
430
-
 ATP pH 7.5, 37°C, wild-type enzyme Homo sapiens
440
-
 ATP pH 7.5, 37°C, mutant enzyme E245S Homo sapiens
610
-
 ATP pH 7.5, 37°C, mutant enzyme Q242S Homo sapiens