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Literature summary extracted from
- Structural basis of fatty acid substrate binding to cyclooxygenase-2 (2010), J. Biol. Chem., 285, 22152-22163.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.99.1 | expression of His-tagged wild-type and mutant COX-2 proteins | Mus musculus |
Crystallization (Commentary)
| EC Number | Crystallization (Comment) | Organism |
|---|---|---|
| 1.14.99.1 | arachidonic acid, eicosapentaenoic acid, and docosahexaenoic acid bound to Co3+-protoporphyrin IX-reconstituted recombinant His-tagged isozyme COX-2 mutant N580A, 0.003 ml of 3 mg/ml protein in 25 mM Tris, pH 8.0, 150 mM NaCl, and 0.53% w/v beta-octylglucoside is mixed with 0.003 ml of reservoir solution containing 23-34% polyacrylic acid 5100, 100 mM HEPES, pH 7.5, 20 mM MgCl2, and 0.6% w/v beta-octylglucoside, equilibration over reservoir solution without beta-octylglucoside, 23°C, 3 days to 4 weeks, X-ray diffraction structure determination and analysis at 2.1 A, 2.4 A, and 2.65 A resolution, respectively | Mus musculus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 1.14.99.1 | L531A | site-directed mutagenesis, the mutant shows reduced Vmax and Km with arachidonate compared to the wild-type COX-2 | Mus musculus |
| 1.14.99.1 | L531F | site-directed mutagenesis, the mutant shows reduced Vmax and Km with arachidonate compared to the wild-type COX-2 | Mus musculus |
| 1.14.99.1 | L531P | site-directed mutagenesis, the mutant shows reduced Vmax and Km with arachidonate compared to the wild-type COX-2 | Mus musculus |
| 1.14.99.1 | L531T | site-directed mutagenesis, the mutant shows reduced Vmax and Km with arachidonate compared to the wild-type COX-2 | Mus musculus |
| 1.14.99.1 | N580A | site-directed mutagenesis, crystal structure determination with bound substrates, overview | Mus musculus |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.99.1 | additional information | - |
additional information | Michaelis-Menten kinetics, overview | Mus musculus | |
| 1.14.99.1 | 0.00295 | - |
arachidonate | pH 8.0, 25°C, COX-2 mutant L531P | Mus musculus |  |
| 1.14.99.1 | 0.00365 | - |
arachidonate | pH 8.0, 25°C, COX-2 mutant L531A | Mus musculus | |
| 1.14.99.1 | 0.00514 | - |
arachidonate | pH 8.0, 25°C, COX-2 wild-type COX-2 and mutant N580A | Mus musculus | |
| 1.14.99.1 | 0.00945 | - |
(5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
| 1.14.99.1 | 0.0368 | - |
(4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.14.99.1 | membrane | associated | Mus musculus | 16020 | - |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.1 | arachidonate + AH2 + O2 | Mus musculus | - |
prostaglandin H2 + A + H2O | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.99.1 | Mus musculus | Q05769 | - |
- |
Posttranslational Modification
| EC Number | Posttranslational Modification | Comment | Organism |
|---|---|---|---|
| 1.14.99.1 | glycoprotein | - |
Mus musculus |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.14.99.1 | recombinant His-tagged wild-type and mutant COX-2 proteins by nickel affinity chromatography and gel filtration | Mus musculus |
Renatured (Commentary)
| EC Number | Renatured (Comment) | Organism |
|---|---|---|
| 1.14.99.1 | the purified recombinant apoenzyme is reconstituted with Co3+-protoporphyrin IX and fatty acid substrate to generate the appropriate enzyme-substrate complexes for X-ray crystallographic analyses | Mus musculus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.1 | (4Z,7Z,10Z,13Z,16Z,19Z)-docosahexaenoic acid + AH2 + O2 | the substrate is bound in the cyclooxygenase channel of COX-2, binding structure, overview. The carboxylate of docosahexaenoate interacts with Arg120 and Tyr355 at the base of the channel and the omega-end abuts the side chain of Ile377 near Gly533 in the hydrophobic groove above Ser530 | Mus musculus | ? + A + H2O | - |
? | |
| 1.14.99.1 | (5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic acid + AH2 + O2 | substrate binding structure and nonproductive conformation, overview | Mus musculus | ? + A + H2O | - |
? | |
| 1.14.99.1 | arachidonate + AH2 + O2 | - |
Mus musculus | prostaglandin H2 + A + H2O | - |
? | |
| 1.14.99.1 | arachidonate + AH2 + O2 | substrate binding structure and nonproductive conformation, overview | Mus musculus | prostaglandin H2 + A + H2O | - |
? | |
| 1.14.99.1 | additional information | although arachidonic acid is the preferred substrate, other fatty acids are oxygenated by the isozymes with varying efficiencies. The substrates bind in different conformations in each monomer constituting the homodimer in their respective structures such that one monomer exhibits nonproductive binding and the other productive binding of the substrate in the cyclooxygenase channel, Arg120 and Leu531 play a role, overview | Mus musculus | ? | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.14.99.1 | homodimer | - |
Mus musculus |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.14.99.1 | COX-1 | - |
Mus musculus |
| 1.14.99.1 | COX-2 | - |
Mus musculus |
| 1.14.99.1 | cyclooxygenase-2 | - |
Mus musculus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.1 | 25 | - |
assay at | Mus musculus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.99.1 | 3.45 | - |
(4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
| 1.14.99.1 | 8.7 | - |
(5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
| 1.14.99.1 | 27 | - |
arachidonate | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.1 | 8 | - |
assay at | Mus musculus |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.14.99.1 | heme | - |
Mus musculus | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.14.99.1 | 9 | - |
(4Z,7Z,10Z,13Z,16Z,19Z)-Docosahexaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
| 1.14.99.1 | 920 | - |
(5Z,8Z,11Z,14Z,17Z)-eicosapentaenoic acid | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
| 1.14.99.1 | 5240 | - |
arachidonate | pH 8.0, 25°C, COX-2 mutant N580A | Mus musculus | |
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Release 2023.1 (January 2023)
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