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Literature summary extracted from

  • Rishavy, M.A.; Hallgren, K.W.; Yakubenko, A.V.; Shtofman, R.L.; Runge, K.W.; Berkner, K.L.
    Bronsted analysis reveals Lys218 as the carboxylase active site base that deprotonates vitamin K hydroquinone to initiate vitamin K-dependent protein carboxylation (2006), Biochemistry, 45, 13239-13248.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.1.90 mutational analysis is performed using an expression system lacking endogenous carboxylase. Construction of baculo viruses containing FLAG-tagged carboxylase mutants and expression in infected SF21 cells. Expresses the 758 amino acid human VKD carboxylase bearing a C-terminal extension of AAADYKDDDDK, where the last eight amino acids are the FLAG epitope Homo sapiens

Protein Variants

EC Number Protein Variants Comment Organism
1.14.99.20 H160A 94% of wild-type activity Leptospira interrogans
1.14.99.20 H287A 163% of wild-type activity Leptospira interrogans
1.14.99.20 H381A 142% of wild-type activity Leptospira interrogans
1.14.99.20 K218A no residual activity, but exogenous amines restore activity and its degree depends on the basicity of the amine Leptospira interrogans
4.1.1.90 H160A His to Ala mutants all show full epoxidase activity Homo sapiens
4.1.1.90 H287A His to Ala mutants all show full epoxidase activity Homo sapiens
4.1.1.90 H381A His to Ala mutants all show full epoxidase activity Homo sapiens
4.1.1.90 K218A K218A activity is not detectable. The addition of exogenous amines restores K218A activity while having little effect on wild type carboxylase Homo sapiens

Organism

EC Number Organism UniProt Comment Textmining
1.14.99.20 Leptospira interrogans
-
-
-
4.1.1.90 Homo sapiens
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.1.90 affinity chromatography Homo sapiens

Reaction

EC Number Reaction Comment Organism Reaction ID
1.14.99.20 phylloquinone + reduced acceptor + O2 = 2,3-epoxyphylloquinone + acceptor + H2O K218 is the essential base that deprotonates AH2 to initiate he reaction Leptospira interrogans
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.1.90 0.0000026
-
 H160A mutant Homo sapiens
4.1.1.90 0.0000027
-
 wild type Homo sapiens
4.1.1.90 0.0000045
-
 H381A mutant Homo sapiens

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.1.90 L-glutamate + CO2 + O2 + vitamin K hydroquinone
-
 Homo sapiens gamma-carboxy L-glutamate + vitamin K epoxide + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.1.1.90 vitamin K-dependent carboxylase
-
 Homo sapiens
4.1.1.90 VKD carboxylase
-
 Homo sapiens

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.1.1.90 7.5
-
-
 Homo sapiens
4.1.1.90 8.5
-
 assay at Homo sapiens

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.1.1.90 5.5 8.5
-
 Homo sapiens

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.90 vitamin K
-
 Homo sapiens