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Literature summary extracted from
- Profactor IX propeptide and glutamate substrate binding sites on the vitamin K-dependent carboxylase identified by site-directed mutagenesis (1996), J. Biol. Chem., 271, 17837-17844.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 1.14.99.20 | wild-type and mutants of bovine enzyme species expressed in chinese hamster ovary cells CHO-Dukx-B11 | Bos taurus |
| 4.1.1.90 | expressed in Chinese hamster ovary cells with an immunodetectable octapeptide inserted at the amino-terminal ends | Bos taurus |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 4.1.1.90 | E567A/K569A | CBX567/568 | Bos taurus |
| 4.1.1.90 | E612A/D614A | CBX612/614 | Bos taurus |
| 4.1.1.90 | H177A/H178A | CBX177/178 | Bos taurus |
| 4.1.1.90 | H678A/E679A/R680A | CBX678/679/680 | Bos taurus |
| 4.1.1.90 | K217A/K218A | inactive, CBX217/218 | Bos taurus |
| 4.1.1.90 | K346A/R347A | CBX346/347 | Bos taurus |
| 4.1.1.90 | K438A/D439A/H440A | CBX438/439/440 | Bos taurus |
| 4.1.1.90 | K622A/E623A/K624A | CBX622/623/624 | Bos taurus |
| 4.1.1.90 | additional information | R234A/H235A mutant, R406A/H408A mutant, and R513A/K515A mutant are more susceptible to inhibition by vitamin KH2 than wild type enzyme. R234A/H235A mutant and R406A/H408A mutant exhibit maximal activity at 111 mM vitamin KH2 and R513A/K515A mutant at 56 mM vitamin KH2 | Bos taurus |
| 4.1.1.90 | R189A/K190A/R191A | CBX189/190/191 | Bos taurus |
| 4.1.1.90 | R234A/H235A | vitamin K epoxidase activities are reduced in parallel with the carboxylase activities. Showed defects in the propeptide binding site. Slightly faster mobility than wild-type FLAG-CBX. CBX234/235 | Bos taurus |
| 4.1.1.90 | R359A/H360A/K361A | vitamin K epoxidase activities are reduced in parallel with the carboxylase activities. Showed defects in the propeptide binding site. CBX359/360/361 | Bos taurus |
| 4.1.1.90 | R406A/H408A | vitamin K epoxidase activities are reduced in parallel with the carboxylase activities. Showed defects in the propeptide binding site. CBX406/408 | Bos taurus |
| 4.1.1.90 | R416A/D417A | CBX416/417 | Bos taurus |
| 4.1.1.90 | R513A/K515A | vitamin K epoxidase activities are reduced in parallel with the carboxylase activities. The show normal affinity for the propeptide, FLEEL, proPT28, and vitamin K hydroquinone but exhibited a low catalytic rate for carboxylation. CBX513/515 | Bos taurus |
| 4.1.1.90 | R661A/R662A | CBX622/623/624 | Bos taurus |
| 4.1.1.90 | R671A/R672A/R673A | CBX671/672/673 | Bos taurus |
| 4.1.1.90 | R687A/K688A | CBX687/688 | Bos taurus |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 4.1.1.90 | vitamin K | carboxylation of FLEEL by bovine liver carboxylase is inhibited by high concentrations of vitamin KH2. vitamin K (up to 400 mM) and vitamin K epoxide (up to 1 mM) are not inhibitory. R234A/H235A mutant, R406A/H408A mutant, and R513A/K515A mutant are more susceptible to inhibition by vitamin KH2 than wild type enzyme. R234A/H235A mutant and R406A/H408A mutant exhibit maximal activity at 111 mM vitamin KH2 and R513A/K515A mutant at 56 mM vitamin KH2 | Bos taurus |  |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.1.1.90 | 0.0023 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.0038 | - |
vitamin KH2 | proPT28, R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.004 | - |
vitamin KH2 | proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.0041 | - |
YVFLDHQDADANLILNRPKR | R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 0.0052 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.007 | - |
vitamin KH2 | proPT28, R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.0081 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.0123 | - |
vitamin KH2 | proPT28, R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.0144 | - |
YVFLDHQDADANLILNRPKR | R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.032 | - |
vitamin KH2 | FLEEL, R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.043 | - |
vitamin KH2 | FLEEL, R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 0.053 | - |
vitamin KH2 | FLEEL, R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.074 | - |
vitamin KH2 | FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.097 | - |
vitamin KH2 | FLEEL, R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.78 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 0.78 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 0.91 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 0.92 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 1 | - |
FLEEL | wild type | Bos taurus | |
| 4.1.1.90 | 1.01 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, proIX18 concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 1.1 | - |
FLEEL | FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 1.23 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 1.25 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 1.3 | - |
FLEEL | R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 1.5 | - |
FLEEL | for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 1.5 | - |
FLEEL | R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 1.54 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 1.6 | - |
FLEEL | R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 1.78 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 1.9 | - |
FLEEL | R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 2.01 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
| 4.1.1.90 | 2.03 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
| 4.1.1.90 | 2.06 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 4.1.1.90 | endoplasmic reticulum membrane | - |
Bos taurus | 5789 | - |
| 4.1.1.90 | microsome | - |
Bos taurus | - |
- |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 4.1.1.90 | 99000 | - |
SDS-PAGE, Western Blot. CHO cells transfected with the cDNA for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.20 | phylloquinone + electron donor + O2 | Bos taurus | - |
2,3-epoxyphylloquinone + oxidized electron + H2O | - |
? | |
| 4.1.1.90 | L-glutamate + CO2 + O2 + vitamin K hydroquinone | Bos taurus | - |
gamma-carboxy L-glutamate + vitamin K epoxide + H2O | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.14.99.20 | Bos taurus | - |
- |
- |
| 4.1.1.90 | Bos taurus | Q07175 | - |
- |
Reaction
| EC Number | Reaction | Comment | Organism | Reaction ID |
|---|---|---|---|---|
| 1.14.99.20 | phylloquinone + reduced acceptor + O2 = 2,3-epoxyphylloquinone + acceptor + H2O | enzyme has two distinct functions: vitamin K epoxidation and gamma-glutamyl carboxylation | Bos taurus |
Source Tissue
| EC Number | Source Tissue | Comment | Organism | Textmining |
|---|---|---|---|---|
| 1.14.99.20 | liver | - |
Bos taurus | - |
| 4.1.1.90 | liver | - |
Bos taurus | - |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.20 | additional information | - |
- |
Bos taurus |
Storage Stability
| EC Number | Storage Stability | Organism |
|---|---|---|
| 1.14.99.20 | -80°C, 20% glycerol | Bos taurus |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.14.99.20 | phylloquinone + electron donor + O2 | - |
Bos taurus | 2,3-epoxyphylloquinone + oxidized electron donor + H2O | - |
? | |
| 1.14.99.20 | phylloquinone + electron donor + O2 | - |
Bos taurus | 2,3-epoxyphylloquinone + oxidized electron + H2O | - |
? | |
| 4.1.1.90 | FLEEL + CO2 + O2 + vitamin KH2 | - |
Bos taurus | ? + vitamin K epoxide + H2O | - |
? | |
| 4.1.1.90 | L-glutamate + CO2 + O2 + vitamin K hydroquinone | - |
Bos taurus | gamma-carboxy L-glutamate + vitamin K epoxide + H2O | - |
? | |
| 4.1.1.90 | YVFLDHQDADANLILNRPKR + CO2 + O2 + vitamin KH2 | - |
Bos taurus | ? + vitamin K epoxide + H2O | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 4.1.1.90 | ? | x * 99000, SDS-PAGE | Bos taurus |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 4.1.1.90 | gamma-glutamyl carboxylase | - |
Bos taurus |
| 4.1.1.90 | glutamate carboxylase | - |
Bos taurus |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 1.14.99.20 | 25 | - |
enzyme assay | Bos taurus |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 4.1.1.90 | 0.002 | - |
YVFLDHQDADANLILNRPKR | R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 0.01 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
| 4.1.1.90 | 0.015 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
| 4.1.1.90 | 0.018 | - |
vitamin KH2 | proPT28, R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.02 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.02 | - |
vitamin KH2 | proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.025 | - |
vitamin KH2 | proPT28, R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.033 | - |
vitamin KH2 | proPT28, R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.042 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.056 | - |
YVFLDHQDADANLILNRPKR | R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.059 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM | Bos taurus | |
| 4.1.1.90 | 0.06 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.07 | - |
FLEEL | R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.1 | - |
vitamin KH2 | FLEEL, R513A/K515A mutant | Bos taurus | |
| 4.1.1.90 | 0.11 | - |
vitamin KH2 | FLEEL, R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 0.153 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 0.175 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 0.19 | - |
FLEEL | R359A/H360A/K361A mutant | Bos taurus | |
| 4.1.1.90 | 0.324 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM | Bos taurus | |
| 4.1.1.90 | 0.347 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 0.391 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 0.465 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM | Bos taurus | |
| 4.1.1.90 | 0.488 | - |
YVFLDHQDADANLILNRPKR | FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 0.56 | - |
vitamin KH2 | FLEEL, R406A/H408A mutant | Bos taurus | |
| 4.1.1.90 | 0.56 | - |
YVFLDHQDADANLILNRPKR | R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 0.6 | - |
vitamin KH2 | FLEEL, R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.6 | - |
FLEEL | for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.64 | - |
FLEEL | R234A/H235A mutant | Bos taurus | |
| 4.1.1.90 | 0.645 | - |
YVFLDHQDADANLILNRPKR | R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM | Bos taurus | |
| 4.1.1.90 | 0.66 | - |
vitamin KH2 | FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.7 | - |
FLEEL | wild type | Bos taurus | |
| 4.1.1.90 | 0.7 | - |
FLEEL | FLAG-vitamin K-dependent gamma-glutamyl carboxylase | Bos taurus | |
| 4.1.1.90 | 0.72 | - |
FLEEL | R406A/H408A mutant | Bos taurus | |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 4.1.1.90 | vitamin K | - |
Bos taurus | |
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