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Literature summary for 4.1.1.90 extracted from

  • Sugiura, I.; Furie, B.; Walsh, C.T.; Furie, B.C.
    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.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Chinese hamster ovary cells with an immunodetectable octapeptide inserted at the amino-terminal ends Bos taurus

Protein Variants

Protein Variants Comment Organism
E567A/K569A CBX567/568 Bos taurus
E612A/D614A CBX612/614 Bos taurus
H177A/H178A CBX177/178 Bos taurus
H678A/E679A/R680A CBX678/679/680 Bos taurus
K217A/K218A inactive, CBX217/218 Bos taurus
K346A/R347A CBX346/347 Bos taurus
K438A/D439A/H440A CBX438/439/440 Bos taurus
K622A/E623A/K624A CBX622/623/624 Bos taurus
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
R189A/K190A/R191A CBX189/190/191 Bos taurus
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
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
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
R416A/D417A CBX416/417 Bos taurus
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
R661A/R662A CBX622/623/624 Bos taurus
R671A/R672A/R673A CBX671/672/673 Bos taurus
R687A/K688A CBX687/688 Bos taurus

Inhibitors

Inhibitors Comment Organism Structure
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]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0023
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.0038
-
vitamin KH2 proPT28, R406A/H408A mutant Bos taurus
0.004
-
vitamin KH2 proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.0041
-
YVFLDHQDADANLILNRPKR R359A/H360A/K361A mutant Bos taurus
0.0052
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant Bos taurus
0.007
-
vitamin KH2 proPT28, R234A/H235A mutant Bos taurus
0.0081
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant Bos taurus
0.0123
-
vitamin KH2 proPT28, R513A/K515A mutant Bos taurus
0.0144
-
YVFLDHQDADANLILNRPKR R513A/K515A mutant Bos taurus
0.032
-
vitamin KH2 FLEEL, R513A/K515A mutant Bos taurus
0.043
-
vitamin KH2 FLEEL, R359A/H360A/K361A mutant Bos taurus
0.053
-
vitamin KH2 FLEEL, R406A/H408A mutant Bos taurus
0.074
-
vitamin KH2 FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.097
-
vitamin KH2 FLEEL, R234A/H235A mutant Bos taurus
0.78
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
0.78
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM Bos taurus
0.91
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
0.92
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM Bos taurus
1
-
FLEEL wild type Bos taurus
1.01
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, proIX18 concentration 160 microM Bos taurus
1.1
-
FLEEL FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
1.23
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
1.25
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
1.3
-
FLEEL R234A/H235A mutant Bos taurus
1.5
-
FLEEL for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
1.5
-
FLEEL R359A/H360A/K361A mutant Bos taurus
1.54
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
1.6
-
FLEEL R406A/H408A mutant Bos taurus
1.78
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
1.9
-
FLEEL R513A/K515A mutant Bos taurus
2.01
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus
2.03
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus
2.06
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus

Localization

Localization Comment Organism GeneOntology No. Textmining
endoplasmic reticulum membrane
-
Bos taurus 5789
-
microsome
-
Bos taurus
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
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)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-glutamate + CO2 + O2 + vitamin K hydroquinone Bos taurus
-
gamma-carboxy L-glutamate + vitamin K epoxide + H2O
-
?

Organism

Organism UniProt Comment Textmining
Bos taurus Q07175
-
-

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
Bos taurus
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
FLEEL + CO2 + O2 + vitamin KH2
-
Bos taurus ? + vitamin K epoxide + H2O
-
?
L-glutamate + CO2 + O2 + vitamin K hydroquinone
-
Bos taurus gamma-carboxy L-glutamate + vitamin K epoxide + H2O
-
?
YVFLDHQDADANLILNRPKR + CO2 + O2 + vitamin KH2
-
Bos taurus ? + vitamin K epoxide + H2O
-
?

Subunits

Subunits Comment Organism
? x * 99000, SDS-PAGE Bos taurus

Synonyms

Synonyms Comment Organism
gamma-glutamyl carboxylase
-
Bos taurus
glutamate carboxylase
-
Bos taurus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.002
-
YVFLDHQDADANLILNRPKR R359A/H360A/K361A mutant Bos taurus
0.01
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus
0.015
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus
0.018
-
vitamin KH2 proPT28, R513A/K515A mutant Bos taurus
0.02
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.02
-
vitamin KH2 proPT28, FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.025
-
vitamin KH2 proPT28, R406A/H408A mutant Bos taurus
0.033
-
vitamin KH2 proPT28, R234A/H235A mutant Bos taurus
0.042
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant Bos taurus
0.056
-
YVFLDHQDADANLILNRPKR R513A/K515A mutant Bos taurus
0.059
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0 microM Bos taurus
0.06
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant Bos taurus
0.07
-
FLEEL R513A/K515A mutant Bos taurus
0.1
-
vitamin KH2 FLEEL, R513A/K515A mutant Bos taurus
0.11
-
vitamin KH2 FLEEL, R359A/H360A/K361A mutant Bos taurus
0.153
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
0.175
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
0.19
-
FLEEL R359A/H360A/K361A mutant Bos taurus
0.324
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 0.16 microM Bos taurus
0.347
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
0.391
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
0.465
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 1.6 microM Bos taurus
0.488
-
YVFLDHQDADANLILNRPKR FLAG-vitamin K-dependent gamma-glutamyl carboxylase, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM Bos taurus
0.56
-
vitamin KH2 FLEEL, R406A/H408A mutant Bos taurus
0.56
-
YVFLDHQDADANLILNRPKR R406A/H408A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM Bos taurus
0.6
-
vitamin KH2 FLEEL, R234A/H235A mutant Bos taurus
0.6
-
FLEEL for wild type FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.64
-
FLEEL R234A/H235A mutant Bos taurus
0.645
-
YVFLDHQDADANLILNRPKR R234A/H235A mutant, peptides comprising residue YVFLDHQDADANLILNRPKR concentration 160 microM Bos taurus
0.66
-
vitamin KH2 FLEEL, FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.7
-
FLEEL wild type Bos taurus
0.7
-
FLEEL FLAG-vitamin K-dependent gamma-glutamyl carboxylase Bos taurus
0.72
-
FLEEL R406A/H408A mutant Bos taurus

Cofactor

Cofactor Comment Organism Structure
vitamin K
-
Bos taurus