Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli BL21 and Rosetta cells. Expression in the baculovirus expression system produced only insoluble, aggregated protein | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
in complex with methyl arachidonyl fluorophosphonate, hanging drop vapor diffusion method, using 6-10% (w/v) PEGMME 5000, 100 mM Na-MES pH 6.0, 0.2% (w/v) glucopyranoside, at 22°C | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
L167Q/L171Q | the mutant shows increased catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L167Q/L174Q | the mutant shows increased catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L169S/L176S | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate and increased catalytic efficiency with umbelliferyl arachidonate compared to the wild type enzyme | Homo sapiens |
L169S/L176S | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L169S/L176S/K160A | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L169S/L176S/K165A | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L169S/L176S/K226A | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L169S/L176S/K36A | the mutant shows about wild type catalytic efficiency with 4-methylumbelliferyl butyrate | Homo sapiens |
L169S/L176S/K36A/K226A | the mutant shows reduced catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
L171Q | the mutant shows about wild type catalytic efficiency with 4-methylumbelliferyl butyrate | Homo sapiens |
L171Q/L174Q | the mutant shows increased catalytic efficiency with 4-methylumbelliferyl butyrate compared to the wild type enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
methyl arachidonyl fluorophosphonate | irreversible active-site inhibitor of monoglyceride lipase | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.084 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.084 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.089 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.09 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K160A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.105 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.11 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.123 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.124 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.136 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.137 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K165A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.162 | - |
4-Methylumbelliferyl butyrate | wild type enzyme, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | - |
Homo sapiens | 16020 | - |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-arachidonoylglycerol + H2O | Homo sapiens | - |
glycerol + arachidonic acid | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q99685 | - |
- |
Purification (Comment) | Organism |
---|---|
His-Trap column chromatography and Superdex 200 gel filtration | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2-arachidonoylglycerol + H2O | - |
Homo sapiens | glycerol + arachidonic acid | - |
? | |
4-methylumbelliferyl butyrate + H2O | - |
Homo sapiens | 4-methylumbelliferol + butyrate | - |
? | |
umbelliferyl arachidonate + H2O | - |
Homo sapiens | umbelliferol + arachidonic acid | - |
? |
Synonyms | Comment | Organism |
---|---|---|
MGL | - |
Homo sapiens |
monoglyceride lipase | - |
Homo sapiens |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
58 | - |
the midpoint of the melting transition is calculated to be 58°C for wild type enzyme | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.45 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K165A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.5 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K160A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.5 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.63 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.78 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.85 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.85 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
0.98 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
1.13 | - |
4-Methylumbelliferyl butyrate | wild type enzyme, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
1.17 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.5 | - |
umbelliferyl arachidonate | wild type enzyme, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
1.7 | - |
umbelliferyl arachidonate | mutant enzyme L169S/L176S, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
3.3 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K165A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
4.2 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
5.5 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K160A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
5.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
5.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K226A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
6.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L169S/L176S/K36A, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
7 | - |
4-Methylumbelliferyl butyrate | wild type enzyme, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
8.7 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L171Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
11.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L171Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens | |
13.8 | - |
4-Methylumbelliferyl butyrate | mutant enzyme L167Q/L174Q, in 20 mM PIPES, pH 7.0, and 150 mM NaCl at 37°C | Homo sapiens |