Cloned (Comment) | Organism |
---|---|
expression of mutant enzyme S65T, S65A and E66D by using an expression system with baculovirus/insect cells, expression in COS cells | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
E66D | KM-value and maximal velocity with 4-methylumbelliferyl alpha-D-galactoside as substrate are similar to the wild-type values. The stability of the mutant enzyme at neutral pH is reduced to 54% of the wild-type enzyme. The intracellular activity in COS1 cells is 68% of the wild-type activity. Intracellular activity in COS1 cells is enhanced to 80% of wild-type activity by cultivation of cells with 1-deoxygalactonojirimycin | Homo sapiens |
S65A | KM-value and maximal velocity with 4-methylumbelliferyl alpha-D-galactoside as substrate are similar to the wild-type values. The stability of the mutant enzyme at neutral pH is reduced to 29% of the wild-type enzyme. The intracellular activity in COS1 cells is 26% of the wild-type activity. Intracellular activity in COS1 cells is enhanced to 44% of wild-type activity by cultivation of cells with 1-deoxygalactonojirimycin | Homo sapiens |
S65T | KM-value and maximal velocity with 4-methylumbelliferyl alpha-D-galactoside as substrate are similar to the wild-type values. The stability of the mutant enzyme at neutral pH is reduced to 4% of the wild-type enzyme. The intracellular activity in COS1 cells is 9% of the wild-type activity. Intracellular activity in COS1 cells is enhanced to 34% of wild-type activity by cultivation of cells with 1-deoxygalactonojirimycin | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
1-deoxygalactonojirimycin | IC50: 63 nM with mutant enzyme S65T, 48 nM with mutant enzyme S65A, 58 nM with mutant enzyme S65D and 59 nM with wild-type enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.3 | - |
4-methylumbelliferyl alpha-D-galactoside | pH 4.6, mutant enzyme E66D | Homo sapiens | |
2.3 | - |
4-methylumbelliferyl alpha-D-galactoside | pH 4.6, wild-type enzyme | Homo sapiens | |
3.4 | - |
4-methylumbelliferyl alpha-D-galactoside | pH 4.6, mutant enzyme S65A | Homo sapiens | |
3.5 | - |
4-methylumbelliferyl alpha-D-galactoside | pH 4.6, mutant enzyme S65T | Homo sapiens |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
46000 | - |
x * 46000, wild-type and mutant enzymes S65T, S65A and E66D, SDS-PAGE | Homo sapiens |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
glycoprotein | - |
Homo sapiens |
Purification (Comment) | Organism |
---|---|
- |
Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl alpha-D-galactoside + H2O | - |
Homo sapiens | 4-methylumbelliferone + alpha-D-galactose | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 46000, wild-type and mutant enzymes S65T, S65A and E66D, SDS-PAGE | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
alpha-Gal A | - |
Homo sapiens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
20 min, wild-type enzyme is stable in the pH-range 4.0-7.0, mutant enzymes S65T, S65A and E66D are stable in the pH-range 4.0-6.0 | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
4.2 | - |
wild-type and mutant enzymes S65T, S65A and S65D | Homo sapiens |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
3.6 | 5.4 | pH 3.6: about 75% of maximal activity, pH 5.4: about 40% of maximal activity, wild-type enzyme and mutant enzymes S65T, S65A and E66D | Homo sapiens |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
4 | 6 | 37°C, 20 min, mutant enzymes S65T, S65A and E66D are stable | Homo sapiens |
4 | 7 | 37°C, 20 min, wild-type enzyme is stable | Homo sapiens |
6.1 | - |
mutant enzyme S65T, more than 50% loss of activity after 20 min above | Homo sapiens |
6.5 | - |
mutant enzyme S65A, more than 50% loss of activity after 20 min above | Homo sapiens |
6.9 | - |
mutant enzyme S66D, more than 50% loss of activity after 20 min above | Homo sapiens |
7 | - |
20 min,mutant enzyme S65T shows 3% residual activity, mutant enzyme S65A shows 24% residual activity, mutant enzyme S66D shows 45% residual activity | Homo sapiens |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.000048 | - |
IC50 48 nM with mutant enzyme S65A | Homo sapiens | 1-deoxygalactonojirimycin | |
0.000058 | - |
IC50 58 nM with mutant enzyme S65D | Homo sapiens | 1-deoxygalactonojirimycin | |
0.000059 | - |
IC50 59 nM with wild-type enzyme | Homo sapiens | 1-deoxygalactonojirimycin | |
0.000063 | - |
IC50: 63 nM with mutant enzyme S65T | Homo sapiens | 1-deoxygalactonojirimycin |