Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Saccharomyces cerevisiae |
Crystallization (Comment) | Organism |
---|---|
sitting drop vapour diffusion method, using 0.17 M ammonium acetate, 0.085 M sodium acetate trihydrate (pH 4.6), 25.5% (w/v) PEG 4000, and 15% (v/v) glycerol | Saccharomyces cerevisiae |
Protein Variants | Comment | Organism |
---|---|---|
C60H/W197I | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
C60K/W197I | the mutation results in a further enhancement of the rates of phosphorylation with paraoxon but does not affect the dephosphorylation of the enzyme | Saccharomyces cerevisiae |
C60Q/W197I | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
C60R/W197I | the mutation results in a further enhancement of the rates of phosphorylation with paraoxon but does not affect the dephosphorylation of the enzyme | Saccharomyces cerevisiae |
C60S | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
C60S/W197I | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
G57H | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
L58H/W197I | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
M162H | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
M162H/C60S/W197I | mutant shows significantly decreased activity | Saccharomyces cerevisiae |
W197I | the substitution enhances SFGH reactivity with paraoxon by more than 1000fold thereby overcoming natural organophosphate resistance, the mutant increases the rate of organophosphate inhibition under pseudo-first-order conditions but does not accelerate organophosphate hydrolysis | Saccharomyces cerevisiae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Hg2+ | susceptible to inhibition by Hg2+ | Saccharomyces cerevisiae | |
methyl paraoxon | - |
Saccharomyces cerevisiae | |
paraoxon | - |
Saccharomyces cerevisiae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.012 | - |
4-nitrophenyl butyrate | mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.013 | - |
4-nitrophenyl acetate | mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.018 | - |
4-nitrophenyl butyrate | mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.019 | - |
4-nitrophenyl butyrate | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.03 | - |
4-nitrophenyl butyrate | mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.04 | - |
4-nitrophenyl butyrate | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.054 | - |
4-nitrophenyl acetate | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.09 | - |
4-nitrophenyl butyrate | wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.1 | - |
4-nitrophenyl acetate | mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.12 | - |
4-nitrophenyl butyrate | mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.13 | - |
4-nitrophenyl butyrate | mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.14 | - |
4-nitrophenyl acetate | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.14 | - |
4-nitrophenyl butyrate | mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.15 | - |
4-nitrophenyl acetate | mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.19 | - |
4-nitrophenyl acetate | mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.19 | - |
4-nitrophenyl butyrate | mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.2 | - |
4-nitrophenyl butyrate | Km less than 0.2 mM, mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.23 | - |
4-nitrophenyl acetate | mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.34 | - |
4-nitrophenyl acetate | mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.4 | - |
4-nitrophenyl acetate | Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.9 | - |
4-nitrophenyl acetate | mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.25 | - |
S-Lactoylglutathione | Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
2.6 | - |
4-nitrophenyl acetate | mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
3 | - |
S-Lactoylglutathione | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
3 | - |
S-Lactoylglutathione | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
33930 | - |
calculated from amino acid sequence | Saccharomyces cerevisiae |
33934 | - |
1 * 33934, calculated from amino acid sequence | Saccharomyces cerevisiae |
67800 | - |
gel filtration | Saccharomyces cerevisiae |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Saccharomyces cerevisiae | P40363 | - |
- |
Oxidation Stability | Organism |
---|---|
the wild type enzyme is sensitive to oxidation | Saccharomyces cerevisiae |
Purification (Comment) | Organism |
---|---|
- |
Saccharomyces cerevisiae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl acetate + H2O | - |
Saccharomyces cerevisiae | 4-nitrophenol + acetate | - |
? | |
4-nitrophenyl butyrate + H2O | - |
Saccharomyces cerevisiae | 4-nitrophenol + butyrate | - |
? | |
paraoxon + H2O | - |
Saccharomyces cerevisiae | diethylphosphate + 4-nitrophenol | - |
? | |
S-formylglutathione + H2O | - |
Saccharomyces cerevisiae | glutathione + formate | - |
? | |
S-lactoylglutathione + H2O | - |
Saccharomyces cerevisiae | glutathione + lactate | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | the wild type enzyme crystallizes as a dimer of dimers with four molecules in the asymmetric unit, X-ray crystallography | Saccharomyces cerevisiae |
monomer | 1 * 33934, calculated from amino acid sequence | Saccharomyces cerevisiae |
Synonyms | Comment | Organism |
---|---|---|
esterase D | - |
Saccharomyces cerevisiae |
SFGH | - |
Saccharomyces cerevisiae |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.003 | - |
4-nitrophenyl butyrate | mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.0055 | - |
4-nitrophenyl acetate | mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.0165 | - |
4-nitrophenyl butyrate | kcat less than 0.0165 s-1, mutant enzyme L58H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.028 | - |
4-nitrophenyl acetate | mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.04 | - |
4-nitrophenyl butyrate | mutant enzyme M162H/C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.078 | - |
4-nitrophenyl butyrate | mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.1 | - |
4-nitrophenyl acetate | mutant enzyme M162H, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.2 | - |
4-nitrophenyl butyrate | wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.32 | - |
4-nitrophenyl acetate | mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.43 | - |
4-nitrophenyl butyrate | mutant enzyme C60R/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.525 | - |
4-nitrophenyl acetate | mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.76 | - |
4-nitrophenyl acetate | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
0.97 | - |
4-nitrophenyl acetate | mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.05 | - |
4-nitrophenyl acetate | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.23 | - |
4-nitrophenyl butyrate | mutant enzyme C60K/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.5 | - |
4-nitrophenyl acetate | mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.53 | - |
4-nitrophenyl acetate | mutant enzyme C60S, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.65 | - |
4-nitrophenyl butyrate | mutant enzyme C60Q/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.8 | - |
4-nitrophenyl acetate | Km above 1.25 mM, wild type enzyme, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.92 | - |
4-nitrophenyl butyrate | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
1.97 | - |
4-nitrophenyl butyrate | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
3.63 | - |
4-nitrophenyl butyrate | mutant enzyme C60H/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
50 | - |
S-Lactoylglutathione | mutant enzyme C60S/W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae | |
116.7 | - |
S-Lactoylglutathione | mutant enzyme W197I, in 0.067 M Na/K phosphate, at pH 7.4 and 22°C | Saccharomyces cerevisiae |