Information on EC 4.1.1.90 - peptidyl-glutamate 4-carboxylase

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The expected taxonomic range for this enzyme is: Coelomata

EC NUMBER
COMMENTARY
4.1.1.90
-
RECOMMENDED NAME
GeneOntology No.
peptidyl-glutamate 4-carboxylase
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
peptidyl-4-carboxyglutamate + 2,3-epoxyphylloquinone + H2O = peptidyl-glutamate + CO2 + O2 + phylloquinone
show the reaction diagram
-
-
-
-
peptidyl-4-carboxyglutamate + 2,3-epoxyphylloquinone + H2O = peptidyl-glutamate + CO2 + O2 + phylloquinone
show the reaction diagram
in the physiological process reaction runs in reversed direction
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxylation
-
-
gamma-carboxylation
-
-
gamma-carboxylation
Q9W0C4
-
PATHWAY
KEGG Link
MetaCyc Link
Biosynthesis of secondary metabolites
-
Ubiquinone and other terpenoid-quinone biosynthesis
-
SYSTEMATIC NAME
IUBMB Comments
peptidyl-glutamate 4-carboxylase (2-methyl-3-phytyl-1,4-naphthoquinone-epoxidizing)
The enzyme can use various vitamin-K derivatives, including menaquinone, but does not contain iron. In the reverse direction the mechanism appears to involve the generation of a strong base by oxygenation of vitamin K. It catalyses the post-translational modification of several proteins of the blood-clotting system. 9--12 glutamate residues are converted to 4-carboxyglutamate (Gla) in a specific domain of the target protein. The 4-pro-S hydrogen of the glutamate residue is removed [5] and there is an inversion of stereochemistry at this position [6].
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
carboxylase
P38435
-
Ci-GGC
Q008V9
-
gamma glutamyl carboxylase
-
-
gamma-carboxylase
-
-
gamma-glutamyl carboxylase
Q07175
-
gamma-glutamyl carboxylase
Q008V9
-
gamma-glutamyl carboxylase
Q8IA33
-
gamma-glutamyl carboxylase
Q9W0C4
-
gamma-glutamyl carboxylase
P38435
-
gamma-glutamyl carboxylase
P38435
polytopic membrane protein
gamma-glutamyl carboxylase
-
-
gamma-glutamyl carboxylase
O88496
-
GGCX
A7YA96
-
GGCX
P38435
-
GGCX
-
-
GGCX
-
-
glutamate carboxylase
Q8IA33
-
glutamate carboxylase
Q9W0C4
-
glutamate carboxylase
-
-
glutamate carboxylase
P38435
-
glutamate carboxylase
O88496
-
matrix gamma-carboxyglutamate protein
Q07175
-
matrix gamma-carboxyglutamate protein
P38435
-
matrix Gla protein
Q07175
-
matrix Gla protein
P38435
-
peptidyl-glutamate 4-carboxylase
-
-
peptidyl-glutamate 4-carboxylase (2-methyl-3-phytyl-1,4-naphthoquinone-epoxidizing)
-
-
two-chain carboxylase
-
carboxylase and epoxidase activities similar to those of one-chain carboxylase
vitamin K carboxylase
A7YA96
-
vitamin K-dependent carboxylase
Q07175
-
vitamin K-dependent carboxylase
-
-
vitamin K-dependent gamma-glutamyl carboxylase
-
-
vitamin K-dependent gamma-glutamylcarboxylase
-
-
VKC
A7YA96
-
VKD carboxylase
-
-
CAS REGISTRY NUMBER
COMMENTARY
81181-72-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
schematic of amino acid homology between human and the Drosophila gamma-glutamyl carboxylase shown
UniProt
Manually annotated by BRENDA team
Sprague-Dawley rat
UniProt
Manually annotated by BRENDA team
Wistar strain male albino rats, vitamin K-deficient or warfarin-treated
UniProt
Manually annotated by BRENDA team
Atlantic salmon
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
malfunction
-
onset of mineralization in Abcc6-/-, Ggcx+/+ mice is delayed until between 3 and 4 months of age, suggesting that the genetic background plays a role in modifying the mineralization process. Mineralization in the Abcc6-/-, Ggcx+/- mice is accelerated in comparison with age-matched Abcc6-/-, Ggcx+/+ mice, with ca. 3fold difference at 3, 4, and 9 months of age
physiological function
-
genetic variation in GGCX influences the gamma carboxylation of vitamin K-dependent proteins, as measured by differences in percent undercarboxylated osteocalcin and proteins induced by vitamin K absence-factor II (PIVKA-II). Heterozygous carriers of GGCX rs10187424 and rs7568458 have significantly lower percent undercarboxylated osteocalcin relative to either homozygous group. No significant differences within GGCX genotype group for plasma phylloquinone. No significant associations of GGCX polymorphisms and PIVKA-II
physiological function
-
GGCX mutations cause coagulation disorder and pseudoxanthoma elasticum. Two GGCX mutations from affected individuals (p. R83W and p.Q374X). GGCX is necessary for activation of both coagulation factors in the liver and matrix gla protein, which, in fully carboxylated form, is able to prevent ectopic mineralization
physiological function
-
genetic variations in GGCX gene contributes to impaired metabolism of warfarin
physiological function
-
functional GGCX mutations in all 15 exons do not occur in most calcium oxalate urolithiasis patients, thus no significant association between the low activity and mutation of GGCX in calcium oxalate urolithiasis
physiological function
-
two novel splice site mutations in the GGCX gene, G to T transversion of the first nucleotide of intron 2 (c. 1358+1G-T) and an A to G transversion of the third nucleotide of intron 11 (c. 10363+3A-G)
physiological function
-
relevance between the defect of GGCX and urolithasis. Decreased activity and production of GGCX between calcium oxalate urolithiasis patients and control patients. GGCX may play an important role in the formation of calcium oxalate. The decreased expression of GGCX may lead to the defect of activity, and may result in the declined ability of calcium oxalate of urinary prothrombin fragment 1
physiological function
A7YA96
VKC maintains vitamin K levels and sustains the blood coagulation cascade
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
4-benzoyl-L-phenylalanine + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
BPA
-
-
?
CDADWVEGYSMEYLSR + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
CDADWVEGYSMEYLSR
-
-
?
CGRPSLEQLAQEVTYA + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
CGRPSLEQLAQEVTYA
-
-
?
conantokin G + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
Q9W0C4
poorly carboxylated
-
-
?
conotoxin epsilon-TxIX + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
-
-
-
-
e-TxIX12 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues 1-12 of e-TxIX
-
-
?
FLEEL + CO2 + O2 + ammonium sulfate
? + vitamin K epoxide + H2O
show the reaction diagram
-
carboxylase activity is measured by 14CO2 incorporation into the synthetic peptide substrate FLEEL
-
-
?
FLEEL + CO2 + O2 + phylloquinone
? + 2,3-epoxyphylloquinone + H2O
show the reaction diagram
-
-
-
-
?
FLEEL + CO2 + O2 + proFIX 19
? + vitamin K epoxide + H2O
show the reaction diagram
-
carboxylase activity is measured by 14CO2 incorporation into the synthetic peptide substrate FLEEL
-
-
?
FLEEL + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
FLEEL + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
Q07175
-
-
-
?
FLEEL + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
Q9W0C4
-
-
-
?
FLEEL + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
pentapeptide substrate FLEEL: Phe-Leu-Glu-Glu-Leu, used for carboxylation activity
-
-
?
FLEEL + CO2 + O2 + vitamin K1 hydroquinone
? + vitamin K1 epoxide + H2O
show the reaction diagram
-
a synthetic peptide substrate, assay for vitamin K–dependent carboxylase activity
-
-
?
FLEEL + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
FLEEL + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-, Q8IA33
-
-
-
?
FLEEL + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
carboxylase activity is measured by 14CO2 incorporation into the synthetic peptide substrate FLEEL
-
-
?
FLEEL + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
Phe-Leu-Glu-Glu-Leu
-
-
?
FLEEV + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
gamma-carboxylated glutamyl containing vitamin K-dependent protein + vitamin K epoxide + H2O
?
show the reaction diagram
-
-
-
-
?
GKDRLTQMKRILKQRGNKARGEEELY + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
Q9W0C4
-
-
-
?
glutamyl containing vitamin K-dependent protein + CO2 + vitamin K hydroquinone + O2
?
show the reaction diagram
-
propeptide binding increases carboxylase affinity for the Glu substrate, and the coordinated binding of the vitamin K–dependent propeptide and Glu substrate increase carboxylase affinity for vitamin K and activity, possibly through a mechanism of substrate-assisted catalysis. The propeptide adjacent to the Gla domain is cleaved subsequently to carboxylation. The carboxylase uses the energy of vitamin K hydroquinone oxygenation to convert glutamyl residues to gamma-carboxylated glutamyl residues in vitamin K–dependent proteins. During carboxylation, the vitamin K hydroquinone cofactor is oxidized to a vitamin K epoxide product. The carboxylase itself is also a vitamin K–dependent protein and carboxylase carboxylation may be important in regulating the overall process of vitamin K–dependent protein carboxylation. All vitamin K–dependent proteins contain multiple glutamyl residues that undergo carboxylation, which is accomplished by a processive mechanism. A single binding event between carboxylase and vitamin K–dependent protein can give rise to all of the glutamyl to gamm-carboxylated glutamyl conversions in the vitamin K–dependent protein. Carboxylation is limited to the glutamyl residue residing within the Gla domainate
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
P38435
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
Q07175
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
O88496
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
Q9W0C4
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-, Q8IA33
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
A7YA96
-
-
-
ir
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
vitamin K epoxide must be recycled to vitamin K hydroquinone by the enzyme epoxide reductase for the reaction to continue
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
the enzyme uses oxygenation of vitamin K hydroquinone to vitamin K epoxide to drive the incorporation of CO2 into glutamyl residues to produce carboxylated glutamate in vitamin K-dependent proteins
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy-L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy-L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy-L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
catalyzes modification of specific glutamic acids to gamma-carboxyglutamic acid in several blood-coagulation proteins
-
-
?
N-(bromoacetyl)-FLEELY + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
osteocalcin + reduced vitamin K + CO2 + O2
? + vitamin K epoxide + H2O
show the reaction diagram
O88496
-
-
-
?
peptidyl-L-glutamate + CO2 + O2 + menaquinone
?
show the reaction diagram
-
-
-
-
?
peptidyl-L-glutamate + CO2 + O2 + phylloquinone
peptidyl-4-carboxy L-glutamate + 2,3-epoxyphylloquinone + H2O
show the reaction diagram
-
-
-
-
?
pro-e-TxIX/12 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -12 to -1 of e-TxIX precursor
-
-
?
pro-e-TxIX/24 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -12 to +12 of e-TxIX precursor
-
-
?
pro-e-TxIX/41 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -29 to +12 of e-TxIX precursor
-
-
?
pro-FIX19 + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
pro-FIX19: peptide comprising residues of human factor IX AVFLDHENANKILNRPKRY
-
-
?
pro-FIX19-16BPA + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
pro-FIX19-16BPA: peptide comprising residues TVBLDHENANKILNRPKRY
-
-
?
proFIX 19 + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
proFIX 19, peptide sequence: TVFLDHENANKILNRPKRY
-
-
-
ProFIX 19-6BPA + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
TVFLDHENANKIBNRPKR
-
-
?
ProFIX 19-7BPA + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
TVFLDHENfiNKBLNRPKR
-
-
?
proFIX/PT28 + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
proFIX/PT28, peptide sequence: TVFLDHENANKILNRPKRANTFLEEVRK
-
-
?
proFIX18 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -18 to -1 of proFactor IX
-
-
?
proFIX19 + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
TVFLDHENANKILNRPKRY
-
-
?
ProFIX19-13BPA + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
TVFLDBENWKILNRPKRY
-
-
?
ProFIX19-16BPA + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
TVBLDHENANKILNRPKRY
-
-
?
proFIX28 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -18 to +10 of proFactor IX
-
-
?
proFIXl9 + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
AVFLDHENANKILNRPKRY
-
-
?
proPT18 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -18 to -1 of proprothrombin. 28-residue peptides based on residues -18 to +10 of human proprothrombin and proFactor IX with Km values of 420 lM, 1.7 microM and 6 microM
-
-
?
proPT28 + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
HVFLAPQQARSLLQRVRRANTFLEEVRK
-
-
?
proPT28 + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
Q07175
proPT28: synthetic peptide is designated by the following nomenclature: pro indicates the presence of the propeptide sequence, PT indicates prothrombin, the protein on which the peptide sequence is based, and 28 indicates the number of amino acid residues in the peptide
-
-
?
proPT28 + CO2 + O2 + vitamin KH2
?
show the reaction diagram
-, Q8IA33
residues -18 to +10 of proprothrombin
-
-
?
proPTl8 + CO2 + O2 + FLEEL + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
HVFLAPQQARSLLQRVRR
-
-
?
TVFLDHENANKILNRPKRANTBLEEVRK + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
carboxylase probe1, TVFLDHENANKILNRPKRANTBLEEVRK as a substrate
-
-
?
TVFLDHENANKILNRPKRYNTBLEEVRK + CO2 + O2 + vitamin K hydroquinone
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
YVFLDHQDADANLILNRPKR + CO2 + O2 + vitamin KH2
? + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy-L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
reaction is essential for the activity of all of the vitamin K-dependent proteins
-
-
?
additional information
?
-
-
an essential posttranslational modification required for the biological activity of a number of proteins, including proteins involved in blood coagulation and its regulation
-
-
-
additional information
?
-
-
cis-isomer of vitamin K1, the 2-desmethyl derivative of phylloquinone, MK-1, or menadione (2 -methyl-1,4-naphthoquinone) have little or no activity
-
-
-
additional information
?
-
-
enzyme accomplishes the post-translational modification required for the activity of all of the vitamin K-dependent proteins
-
-
-
additional information
?
-
Q07175
enzyme catalyzes the posttranslational modification of specific glutamic acid residues to form gamma-carboxygutamic acid residues within the vitamin K-dependent proteins
-
-
-
additional information
?
-
-
enzyme important for gamma-carboxylation of gla-proteins
-
-
-
additional information
?
-
-
enzyme required for the posttranslational modification of vitamin K-dependent proteins
-
-
-
additional information
?
-
Q9W0C4
one of the most distinctive of the extracellular post-translational modifications is the vitamin K-dependent gamma-carboxylation of glutamate residues to give gamma-carboxyglutamate
-
-
-
additional information
?
-
-
uses the oxygenation of vitamin K to convert glutamyl residues to gamma-carboxylated glutamyl residues in vitamin K–dependent proteins
-
-
-
additional information
?
-
-
vitamin K-dependent carboxylation of glutamate to form gamma-carboxyglutamate (Gla) is unusual among known posttranslational modifications in that substrate recognition does not require a specific sequence around the glutamate residues to be modified
-
-
-
additional information
?
-
-
vitamin K-dependent proteins require carboxylation for activity
-
-
-
additional information
?
-
-, Q8IA33
amino-acid sequences of the synthetic substrates and propeptides are shown
-
-
-
additional information
?
-
-
identification of a striking homology between exon 3 in all known matrix Gla proteins and a 24-residue sequence in the bovine and human gamma-glutamyl carboxylases. Alignment of exon 3 of matrix Gla protein with the homologous region of the gamma-glutamyl carboxylase shown
-
-
-
additional information
?
-
-
photolabeling of Q-glutamyl carboxylase with Bpa peptides
-
-
-
additional information
?
-
O88496
separate active sites are required to support vitamin K-dependent epoxide formation and carboxylation. The binding site for vitamin K oxygenase contains an active thiol group
-
-
-
additional information
?
-
-
vitamin K carboxylase specifically interacts with the propeptide region of the precursor forms of vitamin K dependent proteins
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
Q07175
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
peptidyl-L-glutamate + CO2 + O2 + menaquinone
?
show the reaction diagram
-
-
-
-
?
peptidyl-L-glutamate + CO2 + O2 + phylloquinone
peptidyl-4-carboxy L-glutamate + 2,3-epoxyphylloquinone + H2O
show the reaction diagram
-
-
-
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
gamma-carboxy-L-glutamate + vitamin K epoxide + H2O
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
an essential posttranslational modification required for the biological activity of a number of proteins, including proteins involved in blood coagulation and its regulation
-
-
-
additional information
?
-
-
cis-isomer of vitamin K1, the 2-desmethyl derivative of phylloquinone, MK-1, or menadione (2 -methyl-1,4-naphthoquinone) have little or no activity
-
-
-
additional information
?
-
-
enzyme accomplishes the post-translational modification required for the activity of all of the vitamin K-dependent proteins
-
-
-
additional information
?
-
Q07175
enzyme catalyzes the posttranslational modification of specific glutamic acid residues to form gamma-carboxygutamic acid residues within the vitamin K-dependent proteins
-
-
-
additional information
?
-
-
enzyme important for gamma-carboxylation of gla-proteins
-
-
-
additional information
?
-
-
enzyme required for the posttranslational modification of vitamin K-dependent proteins
-
-
-
additional information
?
-
Q9W0C4
one of the most distinctive of the extracellular post-translational modifications is the vitamin K-dependent gamma-carboxylation of glutamate residues to give gamma-carboxyglutamate
-
-
-
additional information
?
-
-
uses the oxygenation of vitamin K to convert glutamyl residues to gamma-carboxylated glutamyl residues in vitamin K–dependent proteins
-
-
-
additional information
?
-
-
vitamin K-dependent carboxylation of glutamate to form gamma-carboxyglutamate (Gla) is unusual among known posttranslational modifications in that substrate recognition does not require a specific sequence around the glutamate residues to be modified
-
-
-
additional information
?
-
-
vitamin K-dependent proteins require carboxylation for activity
-
-
-
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
phylloquinone
-
-
vitamin K
-
the carboxylase uses the energy of vitamin K hydroquinone oxygenation to convert glutamyl residues to gamma-carboxylated glutamyl residues in vitamin K–dependent proteins. During carboxylation, the vitamin K hydroquinone cofactor is oxidized to a vitamin K epoxide product
vitamin K
O88496
aquamephyton
vitamin K
Q9W0C4
phytonadione
vitamin K
-
conversion of glutamic acid to gamma-carboxyglutamic acid is coupled with the oxygenation of KH2 to vitamin K 2,3-epoxide and has been referred to as vitamin K epoxidase activity
vitamin K
-, Q008V9
-
vitamin K
-
carboxylase and soybean seed lipoxygenase share 19.3% identity over a span of 198 amino acids, from residues 468 to 666 of carboxylase. This is interesting because the carboxylase acts as an oxygenase on the cofactor vitamin K-hydroquinone, and the similarity occurs in that region of the carboxylase likely to have enzymatic function
vitamin K
-
the enzyme requires vitamin K as a cofactor for its post-translational modification of glutamic acid residues to gamma-carboxyglutamic acid residues
vitamin K1
-
-
vitamin K1
-, Q8IA33
-
additional information
-
menadione does not work as a cofactor for gamma-glutamylcarboxylase
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
calcium-binding protein
additional information
-
Mn2+, high concentrations significantly decrease or have no effect on Km of a peptide substrate for the enzyme
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2,3,5,6-Tetrachloro-4-pyridinol
-
TCP, anticoagulant action, effective in vitro inhibitor of the carboxylase
2-chloro-3-phytyl-1,4-naphthoquinone
-
chloro-K, effective in vivo antagonist of vitamin K, inhibits the enzyme in an competitive fashion
2-chloro-3-phytyl-1,4-naphthoquinone
-
-
anti-carboxylase antiserum
-
effect of anti-carboxylase antiserum on carboxylase activity: under the conditions employed the carboxylation is inhibited by 80%, with parallel inhibition of CO2 incorporation into FLEEL and proPT28 (synthetic peptide)
-
Boc-(2S,4S)-4-methylglutamic acid-Glu-Val
-
competitive inhibitor, FLEEL as substrate
Boc-Ser(OPO4)-Ser(OPO4)-Leu-OMe
-
inhibits the enzyme apparently competitively with regard to other peptide substrate
bromoacetyl-FLEEL peptide
-
the His6-carboxylase is irreversibly inactivated. Up to 85% of the carboxylase activity is lost over a period of 120 min
CN-
-
enzyme is blocked by mM concentrations of CN-
Cu2+
-
free Cu2+ and Cu2+-complexes inhibit the reaction
deoxycholate
O88496
-
ethanol
-
high concentrations
FFRCK
-
peptide, protease inhibitor
FLEEL
-
high FLEEL peptide substrate concentrations (from 1.2 up to 12 mM) inhibit GGCX activity
-
FPRCK
-
peptide, protease inhibitor
iodoacetic acid
O88496
poor inhibitor
N-ethylmaleimide
O88496
preincubation with vitamin K hydroquinone prevents NEM inhibition of epoxide formation but not of carboxylation
N-ethylmaleimide
-
-
p-chloromercuribenzoate
O88496
97% inhibition with 1.25 mM and at 5 mM the reaction is completely inhibited
p-hydroxymercuribenzoate
-
-
p-hydroxymercuribenzoate
O88496
1 mM, more than 90% inhibition, inhibition is reversed by dithiothreitol
proFIX/PT28 (Bpa +4)
-
presence of proFIX/PT28 (Bpa +4) or its iodinated derivative 56% inactivation is observed
-
proFIX19-16BPA propeptide
-
-
-
protease inhibitor mixture
-
PIC, freshly prepared as a 10x PIC stock containing 20 mM dithiothreitol, 20 mM EDTA, FFRCK (1.25 microg/ml), FPRCK (1.25 microg/ml), leupeptin (5 microg/ml), pepstatin A (7 microg/ml), phenylmethylsulfonyl fluoride (340 microg/ml), aprotinin (20 microg/ml)
-
Sucrose
O88496
-
tetrachloropyridin
-
-
Trypsin
O88496
-
-
TVFLDHENANKILNRPKRANTBLEEVRK
-
the enzyme is photoirradiated on ice at 365 nm with TVFLDHENANKILNRPKRANTBLEEVRK, TVFLDHENANKILNRPKRYNTBLEEVRK and mono [127I]TVFLDHENANKILNRPKRYNTBLEEVRK for various times
TVFLDHENANKILNRPKRYNTBLEEVRK
-
the enzyme is photoirradiated on ice at 365 nm with TVFLDHENANKILNRPKRANTBLEEVRK, TVFLDHENANKILNRPKRYNTBLEEVRK and mono [127I]TVFLDHENANKILNRPKRYNTBLEEVRK for various times. Presence of TVFLDHENANKILNRPKRYNTBLEEVRK or its iodinated derivative 80% inactivation is observed
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
Warfarin
O88496
-
Warfarin
-
-
Methemoglobin
-
-
-
additional information
-
in the presence of high concentrations of propeptide, only minimal carboxylase activity is measurable. Antibodies to the protein inhibit the carboxylase activity in crude preparations
-
additional information
O88496
NADH, dithiothreitol, and ATP deficiency decrease enzyme activity
-
additional information
-
15 min irradiation in the absence of peptide resulted in a 10% inactivation of the carboxylase
-
additional information
-
the carboxylase reaction is inhibited by sulfhydryl-specific reagents
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
ammonium sulfate
-
-
benzoylphenylalanine
-
Bpa, the four Bpa peptides enhance gamma-carboxylation by 1.5-2.3fold, and the rate enhancement profiles are very similar to that of proFIX19, showing that these propeptides are recognized by the carboxylase
-
decarboxylated plasma prothrombin
-
activates the enzyme
-
endogenous precursor
-
activates the enzyme
-
proFIX 19
-
-
-
proFIX19
-
enhances gamma-carboxylation of the synthetic FLEEL peptide by 2.2-2.3fold
-
proPT18
-
enhances gamma-carboxylation of the synthetic FLEEL peptide by 2.2-2.3fold
-
FLEEL
-
the enzyme activity increases with increasing FLEEL peptide concentration up to 1.2 mM
-
additional information
-
enzyme activity increases 2-3fold by a vitamin K deficiency or Warfarin treatment
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.0017
-
28-residue peptides based on residues -18 to +10 of human proprothrombin
-, Q8IA33
pH 7.0
-
0.3
-
CO2
-
pH 7.4
0.565
-
conotoxin epsilon-TxIX
-, Q8IA33
pH 7.0
-
0.42
-
FLEEL
-, Q8IA33
pH 7.0
-
0.54
-
FLEEL
-
wild type, pH 7.4, 20°C
-
1
-
FLEEL
-
wild type
-
1.1
-
FLEEL
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
1.3
-
FLEEL
-
R234A/H235A mutant
-
1.5
-
FLEEL
-
for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase; R359A/H360A/K361A mutant
-
1.5
-
FLEEL
-
proPTl8 propeptide
-
1.5
-
FLEEL
-
wild-type carboxylase in bovine liver microsomes
-
1.6
-
FLEEL
-
R406A/H408A mutant
-
1.6
-
FLEEL
-
proFIX19 propeptide
-
1.6
-
FLEEL
-
pH 7.4
-
1.6
-
FLEEL
-
recombinant His6-carboxylase present in Sf9 cell microsomes
-
1.8
-
FLEEL
-
proFIX 19-6BPA propeptide
-
1.9
-
FLEEL
-
R513A/K515A mutant
-
1.9
-
FLEEL
-
proFIX19-16BPA propeptide
-
2
-
FLEEL
-
proFIX19-13BPA propeptide; proFIX19-7BPApropeptide
-
6.49
-
FLEEL
-
L394R, pH 7.4, 20°C
-
9
-
FLEEL
-
none propeptide
-
14.8
-
FLEEL
-
Y395A, pH 7.4, 20°C
-
0.075
-
precursor analog containing 12 of the propeptide region
-, Q8IA33
pH 7.0
-
0.074
-
precursor analog containing 29 amino acids of the propeptide region
-, Q8IA33
pH 7.0
-
0.006
-
proFactor IX
-, Q8IA33
pH 7.0
-
0.0068
-
TVFLDHENANKILNRPKRANTBLEEVRK
-
carboxylase probe1, TVFLDHENANKILNRPKRANTBLEEVRK as a substrate
0.052
-
vitamin K1
-, Q8IA33
pH 7.0
0.0038
-
vitamin KH2
-
proPT28, R406A/H408A mutant
-
0.004
-
vitamin KH2
-
proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.007
-
vitamin KH2
-
proPT28, R234A/H235A mutant
-
0.0123
-
vitamin KH2
-
proPT28, R513A/K515A mutant
-
0.032
-
vitamin KH2
-
FLEEL, R513A/K515A mutant
-
0.043
-
vitamin KH2
-
FLEEL, R359A/H360A/K361A mutant
-
0.053
-
vitamin KH2
-
FLEEL, R406A/H408A mutant
-
0.074
-
vitamin KH2
-
FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.097
-
vitamin KH2
-
FLEEL, R234A/H235A mutant
-
0.0023
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.0041
-
YVFLDHQDADANLILNRPKR
-
R359A/H360A/K361A mutant
-
0.0052
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant
-
0.0081
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant
-
0.0144
-
YVFLDHQDADANLILNRPKR
-
R513A/K515A mutant
-
0.78
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM; FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
0.91
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
0.92
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM
-
1.01
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, proIX18 concentration 160 microM
-
1.23
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
1.25
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
1.54
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
1.78
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
2.01
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
2.03
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
2.06
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
24.3
-
FLEEL
-
W399A, pH 7.4, 20°C
-
additional information
-
additional information
-
Km-values measured by hyperbolic weighted least-squares analysis
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
1
-
CO2
-
pH 7.4
0.0019
-
FLEEL
-
Y395A, pH 7.4, 20°C
-
0.0033
-
FLEEL
-
W399A, pH 7.4, 20°C
-
0.07
-
FLEEL
-
R513A/K515A mutant
-
0.11
-
FLEEL
-
L394R, pH 7.4, 20°C
-
0.19
-
FLEEL
-
R359A/H360A/K361A mutant
-
0.21
-
FLEEL
-
wild type, pH 7.4, 20°C
-
0.6
-
FLEEL
-
for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.64
-
FLEEL
-
R234A/H235A mutant
-
0.7
-
FLEEL
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase; wild type
-
0.72
-
FLEEL
-
R406A/H408A mutant
-
1
-
FLEEL
-
wild-type carboxylase in bovine liver microsomes
-
0.018
-
vitamin KH2
-
proPT28, R513A/K515A mutant
-
0.02
-
vitamin KH2
-
proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.025
-
vitamin KH2
-
proPT28, R406A/H408A mutant
-
0.033
-
vitamin KH2
-
proPT28, R234A/H235A mutant
-
0.1
-
vitamin KH2
-
FLEEL, R513A/K515A mutant
-
0.11
-
vitamin KH2
-
FLEEL, R359A/H360A/K361A mutant
-
0.56
-
vitamin KH2
-
FLEEL, R406A/H408A mutant
-
0.6
-
vitamin KH2
-
FLEEL, R234A/H235A mutant
-
0.66
-
vitamin KH2
-
FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.002
-
YVFLDHQDADANLILNRPKR
-
R359A/H360A/K361A mutant
-
0.01
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
0.015
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
0.02
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase
-
0.042
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant
-
0.056
-
YVFLDHQDADANLILNRPKR
-
R513A/K515A mutant
-
0.059
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM
-
0.06
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant
-
0.153
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
0.175
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
0.324
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM
-
0.347
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
0.391
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
0.465
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM
-
0.488
-
YVFLDHQDADANLILNRPKR
-
FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM
-
0.56
-
YVFLDHQDADANLILNRPKR
-
R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM
-
0.645
-
YVFLDHQDADANLILNRPKR
-
R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM
-
1.7
-
FLEEL
-
recombinant His6-carboxylase present in Sf9 cell microsomes
-
additional information
-
additional information
-
kcat values relative to wild type are 51% (L394R), 1% (Y395A), and 2% (W399A), pH 7.4, 20°C
-
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.013
-
Boc-(2S,4S)-4-methylglutamic acid-Glu-Val
-
wild type, pH 7.4, 20°C
1.4
-
Boc-(2S,4S)-4-methylglutamic acid-Glu-Val
-
L394R, pH 7.4, 20°C
0.0174
-
proFIX19-16BPA propeptide
-
competitive inhibition experiments using prom28 substrate
-
2.1
-
Boc-(2S,4S)-4-methylglutamic acid-Glu-Val
-
Y395A, pH 7.4, 20°C
additional information
-
additional information
-
Ki value for Boc-(2S,4S)-4-methylglutamic acid-Glu-Val is above 5 mM for W399A, pH 7.4, 20°C
-
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.0000026
-
-
H160A mutant
0.0000027
-
-
wild type
0.0000045
-
-
H381A mutant
additional information
-
-
specific activities of the partially purified microsomes and the homogeneous vitamin K-dependent carboxylase are 2.77 and 279.6 nmol of vitamin K epoxide per h per mg of protein, a 101fold purification of the vitamin K epoxidase activity from partially purified microsomes
additional information
-
-
propeptide eluate of gel: 608 U/mg, carboxylase units are expressed as 100000 dpm of 14CO2 fixed in the standard assay
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.2
7.4
-
assay at. The activity falls off sharply above pH 8 or below pH 7
7.5
-
-
-
8.5
-
-
assay at
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.5
8.5
-
-
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
coagulation assays
Manually annotated by BRENDA team
-
gamma-carboxylase gene targeting
Manually annotated by BRENDA team
-
vitamin K-dependent carboxylase mRNA expression in early rat embryonic development
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
rat hepatoma cell line
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
adult Sprague-Dawley rat
Manually annotated by BRENDA team
-
preparation of solubilized gamma-carboxylase
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
relatively high GGCX activity compared to other tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
gamma-carboxylase gene targeting
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
-
expression of rat vitamin K-dependent carboxylase in adult and embryonic tissues
Manually annotated by BRENDA team
additional information
-
6-year-old Mexican American male
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
human glutamyl carboxylase spans the membrane at least 5times, with its N-terminus in the cytoplasm and its C-terminus in the lumen of the endoplasmic reticulum
Manually annotated by BRENDA team
-
potential impact of quality control components on carboxylation, which occurs in the endoplasmic reticulum during the secretion of vitamin K–dependent proteins
Manually annotated by BRENDA team
-
transmembrane protein
Manually annotated by BRENDA team
-
amino-terminus of the gamma-glutamyl carboxylase is on the cytoplasmic side of the endoplasmic reticulum, while the carboxylterminus is on the lumenal side
Manually annotated by BRENDA team
-
integral membrane protein
Manually annotated by BRENDA team
-
758 residue integral membrane protein
Manually annotated by BRENDA team
-
microsomal carboxylase activity is compared from cells transfected with pCMV5
-
Manually annotated by BRENDA team
-
from a dicoumarol-treated cow
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
33000
-
-, Q8IA33
cells transfected with the CAT-V5/His plasmid, Western Blot
60000
-
-
proteinase K digestion of the hGC-FLAG reveals a 60 kDa fragment, indicating the lumenal location of the FLAG tag and therefore the carboxyl-terminus of the carboxylase. In contrast, FLAG-hGC does not show a proteinase K–resistant fragment except for the residual undigested full-length carboxylase, which indicates the cytoplasmic location of the FLAG tag and therefore the amino-terminus of the carboxylase
77000
-
-
single major band on SDS gel electrophoresis. The eluted protein contains both stable vitamin K-dependent carboxylase and vitamin K epoxidase activity
85700
-
-
calculated from sequence
87540
-
-
calculated from sequence
94000
-
-
SDS-PAGE
94000
-
-
SDS-PAGE and Western Blot, His6-carboxylase-Ac-FLEEL confirming affinity-purified carboxylase
94000
-
-
SDS-PAGE
94000
-
-
SDS-PAGE
95000
-
-
full-length carboxylase
99000
-
-
SDS-PAGE, Western Blot. CHO cells transfected with the cDNA for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase
130000
-
-, Q8IA33
SDS-PAGE, Sf21 cells transfected with the carboxylase cDNA-containing plasmid, Western Blot
additional information
-
-
determination of disulfide bond formation in purified two-chain carboxylase and P80L and P378L two-chain carboxylases by SDS-PAGE and Western Blot analyses
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 99000, SDS-PAGE
?
-
x * 94000, SDS-PAGE
?
-
x * 94000, SDS-PAGE
heterodimer
A7YA96
-
monomer
-
1 * 77000, SDS-PAGE
monomer
-
-
additional information
-
five transmembrane domains. Transmembrane domain interactions and residue proline 378 are essential for proper structure, especially disulfide bond formation
additional information
-
limited tryptic digestion of the carboxylase yields two disulfide-linked fragments with molecular masses of 30 and 60 kDa, corresponding to the amino and carboxy-terminal part of the gamma-glutamyl carboxylase
additional information
-
binding site in the carboxyl half of the gamma-glutamyl carboxylase. Carboxylase may be cleaved by trypsin into an amino-terminal 30 kDa and a carboxyl-terminal 60 kDa fragment joined by disulfide bond(s), and the propeptide binds to the 60 kDa fragment
additional information
-
disulfide bond between cysteines 99 and 450, five transmembrane domains
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
glycoprotein
-
-
additional information
-
enzyme catalyzes vitamin K-dependent posttranslational modification of glutamate to gamma-carboxyl glutamate
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
enzyme is not very stable at 37°C and lower temperatures are more desirable. At temperatures below 20°C, extended linear rates of incorporation of 14CO2 into exogenous peptide substrates can be observed
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
freeze-thawing, three times, stable
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-70°C, 6 months, stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
affinity chromatography
-
degree of purification is about 7000fold with reference to ammonium sulfate-fractionated microsomal protein from liver. Purification of carboxylase, solubilized microsomes: 281000 cpm/mg/h, flow-through of Affi-FIXQ/S: 200000 cpm/mg/h, bound to Affi-FIXQ/S: 140000000 cpm/mg/h, affinity-purified carboxylase: 1930000000 cpm/mg/h
-
inactivated His6-carboxylase-Ac-FLEEL purified under denaturing conditions by Ni-chelation chromatography followed by preparative polyacrylamide gel electrophoresis
-
partial purification of the enzyme by antibody affinity techniques. Purified 500fold by adsorption to an antiprothrombin column and elution with a dodeca peptide which competes with a prothrombin precursor enzyme recognition site. The purified enzyme is devoid of bound precursors, and has the same ratio of vitamin K epoxidase activity to carboxylase activity as the crude microsomal preparation
-
affinity chromatography
-
difficult, different methods shown
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in baculovirus-infected insect cell. Produced His6-tagged carboxylase as a recombinant protein using a baculovirus expression system
-
expressed in Chinese hamster ovary cells with an immunodetectable octapeptide inserted at the amino-terminal ends
-
His6-tagged bovine liver carboxylase (His6-carboxylase) is produced in insect cells using a baculovirus expression system
-
expression in COS cells or expressed in Sf21 insect cells
-, Q8IA33
expressed in 293 cells
-
expressed in Sf9 cells
-
expression of the cloned cDNA results in an increase in carboxylase activity in microsomes of transfected cells compared to mock-transfected cells
-
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
-
EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
GGCX mRNA in the urolithiasic group is lower than that in the normal control group, which is on average 7.86fold underexpressed in the urolithiasic group compared to the normal control group. Protein expression of GGCX in the urolithiasic group is weaker than that in the control groups
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
E567A/K569A
-
CBX567/568
E612A/D614A
-
CBX612/614
H177A/H178A
-
CBX177/178
H678A/E679A/R680A
-
CBX678/679/680
K217A/K218A
-
inactive, CBX217/218
K346A/R347A
-
CBX346/347
K438A/D439A/H440A
-
CBX438/439/440
R189A/K190A/R191A
-
CBX189/190/191
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
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
R406A/H408A
-
vitamin K epoxidase activities are reduced in parallel with the carboxylase activities. Showed defects in the propeptide binding site. CBX406/408
R416A/D417A
-
CBX416/417
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
R661A/R662A
-
CBX622/623/624
R671A/R672A/R673A
-
CBX671/672/673
R687A/K688A
-
CBX687/688
E373L/Q374L
-
transmembrane domain residues in the C-terminal peptide to test for polar/charge residues
G125L
-
two-chain carboxylase
G128L
-
two-chain carboxylase
G132L
-
two-chain carboxylase
G363L/T367L
-
transmembrane domain residues in the C-terminal peptide to test for polar/charge residues
H160A
-
His to Ala mutants all show full epoxidase activity
H287A
-
His to Ala mutants all show full epoxidase activity
H381A
-
His to Ala mutants all show full epoxidase activity
H404A
-
carboxylases W390A, S398A, and H404A have activities similar to that of wild type
K218A
-
K218A activity is not detectable. The addition of exogenous amines restores K218A activity while having little effect on wild type carboxylase
L128R
-
warfarin resistent mutant
L368/372P
-
mutation to disrupt the transmembrane helix
P378L
-
significantly decreases the disulfide formation in carboxylase
P80L
-
mutation of residue P80, which has activity similar to that of wild-type carboxylase, has a minor effect on disulfide formation
R58G
-
warfarin resistent mutant
S398A
-
carboxylases W390A, S398A, and H404A have activities similar to that of wild type
V29L
-
warfarin resistent mutant
V45A
-
warfarin resistent mutant
W390A
-
carboxylases W390A, S398A, and H404A have activities similar to that of wild type
W399A
-
lower activity than wild type
K622A/E623A/K624A
-
CBX622/623/624
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
L394R
-
natural mutant, certain individuals with combined deficiencies of vitamin K-dependent proteins have a mutation, L394R, in their gamma-glutamyl carboxylase causing impaired glutamate binding
additional information
-
N-terminal carboxylase peptide (residues 1-345) and the C-terminal peptide (345-758) two-chain form (residues 1-345 and residues 346-758) of the vitamin K-dependent gamma-glutamyl carboxylase expressed in Sf9 insect cells. The carboxylase and epoxidase activities similar to those of one-chain carboxylase. The two-chain carboxylase is joined by a disulfide bond
additional information
-
38-BamHI site introduces 2 amino acid residues (glycine and serine) between the hGC fragment and the Lep tag. A 10-amino acid peptide (MDYKDDDDKG), including the FLAG epitope, is introduced to the amino-terminus of the full length of hGC to make FLAG-hGC and a 8-amino acid peptide (DYKDDDDK) is attached to the carboxyl-terminus of the full length of hGC to make hGC-FLAG. The FLAG-tagged hGC cDNA is subcloned into the EcoRI (Escherichia coli RY13) site of the expression vector pCl-neo under control of the cytomegalovirus (CMV) promoter
additional information
-
Y395A propeptide affinity is similar to that of wild type, but those of L394R and W399A are 16-22fold less than that of wild type. Results of kinetic studies with a propeptide-containing substrate are consistent with results of propeptide binding and FLEEL kinetics. Although propeptide and vitamin K binding in some mutants are affected, our data provide compelling evidence that glutamate recognition is the primary function of the conserved region around Leu394
additional information
-
six out of seven patients with Pseudoxanthoma Elasticum habor mutations in the GGCX gene (gamma-glutamyl carboxylase)
additional information
-
GGCX single nucleotide polymorphism rs11676382(but not rs12714145) is a significant predictor of residual warfarin dosing error and is associated with a 6.1% reduction in warfarin dose per G allele
Y395A
-
lower activity than wild type
additional information
-
analysis of a Ggcx+/- intercross reveals a partial developmental block with only 50% of expected Ggcx-/- offspring surviving to term, with the latter animals dying uniformly at birth of massive intra-abdominal hemorrhage. This phenotype closely resembles the partial midembryonic loss and postnatal hemorrhage previously reported for both prothrombin and factor V (F5)–deficient mice. Ggcx-/-, dying uniformly at birth of massive intra-abdominal hemorrhage. Heterozygous mice carrying a null mutation at the gamma-carboxylase (Ggcx) gene exhibit normal development and survival with no evidence of hemorrhage and normal functional activity of the vitamin K–dependent clotting factors IX, X, and prothrombin
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
medicine
-
six out of seven patients with Pseudoxanthoma Elasticum habor mutations in the GGCX gene (gamma-glutamyl carboxylase)
medicine
-
warfarin therapy
medicine
-
multiplexed single nucleotide polymorphism panel (interrogation of the CYP2C9 *2, *3, VKORC1 (-1639G3A), and GGCX (1181T3G) alleles simultaneously) can be successfully used in genotyping of patient blood samples, whereby results can be combined with other clinical parameters in an algorithm for warfarin dosing
medicine
-
single nucleotide polymorphism in the GGCX gene may affect the dose of warfarin and other vitamin K antagonists