γ-Glutamyl carboxylation, a reaction essential for the activity of vitamin K—dependent proteins, requires the concerted actions of γ-glutamyl carboxylase (GGCX), vitamin K 2,3-epoxide reductase complex 1 (VKORC1), and the chaperone calumenin (CALU). We evaluated the contribution of genetic polymorphisms in VKORC1, GGCX, and CALU to interindividual variation in the activities of plasma protein C and protein S. We sequenced these 3 genes in 96 Japanese individuals and genotyped 9 representative single-nucleotide polymorphisms in 3655 Japanese individuals representative of the general population. The mean activity of protein C in women bearing the GG genotype of GGCX 8016G<A (130.8% ± 1.5%, n = 156) was significantly greater (P = .002) than that of individuals with either the AG (126.8% ± 0.7%, n = 728) or the AA (125.4% ± 0.6%, n = 881) genotype, after adjusting for confounding factors. The GGCX 8016G<A change leads to the substitution of Gln for Arg at amino acid residue 325 (Arg325Gln). This effect was comparable to that of a previously defined polymorphism in the protein C promoter. Mean protein S activity was influenced by the VKORC1 3730G<A and CALU 20943T<A genotypes, after adjusting for confounding factors. Thus, polymorphisms in genes involved in the vitamin K—dependent γ-carboxylation reaction influence interindividual variation in the activities of protein C and protein S in the general population.
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Vermeer C. γ-Carboxyglutamate-containing proteins and the vitamin K-dependent carboxylase. Biochem J. 1990;266:625–636.
Furie B, Bouchard BA, Furie BC.Vitamin K-dependent biosynthesis of ³-carboxyglutamic acid. Blood. 1999;93:1798–1808.
Furie B, Furie BC. Molecular basis of vitamin K-dependent γ- carboxylation. Blood. 1990;75:1753–1762.
Stafford DW. The vitamin K cycle. J Thromb Haemost. 2005;3:1873–1878.
Wu SM, Cheung WF, Frazier D, Stafford DW. Cloning and expression of the cDNA for human γ-glutamyl carboxylase. Science. 1991;254:1634–1636.
Tie J, Wu SM, Jin D, Nicchitta CV, Stafford DW.A topological study of the human γ-glutamyl carboxylase. Blood. 2000;96:973–978.
Rost S, Fregin A, Ivaskevicius V, et al. Mutations in VKORC1 cause warfarin resistance and multiple coagulation factor deficiency type 2. Nature. 2004;427:537–541.
Li T, Chang CY, Jin DY, Lin PJ, Khvorova A, Stafford DW. Identification of the gene for vitamin K epoxide reductase. Nature. 2004;427:541–544.
Tie JK, Nicchitta C, von Heijne G, Stafford DW. Membrane topology mapping of vitamin K epoxide reductase by in vitro translation/ cotranslocation. J Biol Chem. 2005;280:16410–16416.
Wajih N, Sane DC, Hutson SM, Wallin R. Engineering of a recombinant vitamin K-dependent γ-carboxylation system with enhanced γ-carboxyglutamic acid forming capacity: evidence for a functional CXXC redox center in the system. J Biol Chem. 2005;280:10540–10547.
Geisen C, Watzka M, Sittinger K, et al. VKORC1 haplotypes and their impact on the inter-individual and inter-ethnical variability of oral anticoagulation. Thromb Haemost. 2005;94:773–779.
Wallin R, Sane DC, Hutson SM. Vitamin K 2,3-epoxide reductase and the vitamin K-dependent γ-carboxylation system. Thromb Res. 2002;108:221–226.
D’Andrea G, D’Ambrosio RL, Di Perna P, et al. A polymorphism in the VKORC1 gene is associated with an interindividual variability in the dose-anticoagulant effect of warfarin. Blood. 2005;105:645–649.
Rieder MJ, Reiner AP, Gage BF, et al. Effect of VKORC1 haplotypes on transcriptional regulation and warfarin dose. N Engl J Med. 2005;352:2285–2293.
Bodin L, Verstuyft C, Tregouet DA, et al. Cytochrome P450 2C9 (CYP2C9) and vitamin K epoxide reductase (VKORC1) genotypes as determinants of acenocoumarol sensitivity. Blood. 2005;106:135–140.
Yuan HY, Chen JJ, Lee MT, et al. A novel functional VKORC1 promoter polymorphism is associated with inter-individual and inter-ethnic differences in warfarin sensitivity. Hum Mol Genet. 2005;14:1745–1751.
Wadelius M, Chen LY, Downes K, et al. Common VKORC1 and GGCX polymorphisms associated with warfarin dose. Pharmacogenomics J. 2005;5:262–270.
Sconce EA,Khan TI,Wynne HA, et al.The impact of CYP2C9 and VKORC1 genetic polymorphism and patient characteristics upon warfarin dose requirements: proposal for a new dosing regimen. Blood. 2005;106:2329–2333.
Mushiroda T, Ohnishi Y, Saito S, et al. Association of VKORC1 and CYP2C9 polymorphisms with warfarin dose requirements in Japanese patients. J Hum Genet. 2006;51:249–253.
Marsh S, King CR, Porche-Sorbet RM, Scott-Horton TJ, Eby CS. Population variation in VKORC1 haplotype structure. J Thromb Haemost. 2006;4:473–474.
Takahashi H, Wilkinson GR, Nutescu EA, et al. Different contributions of polymorphisms in VKORC1 and CYP2C9 to intra- and inter-population differences in maintenance dose of warfarin in Japanese, Caucasians and African-Americans. Pharmacogenet Genomics. 2006;16:101–110.
Yabe D, Nakamura T, Kanazawa N, Tashiro K, Honjo T. Calumenin, a Ca2+-binding protein retained in the endoplasmic reticulum with a novel carboxyl-terminal sequence, HDEF. J Biol Chem. 1997;272:18232–18239.
Wallin R, Hutson SM, Cain D, Sweatt A, Sane DC. A molecular mechanism for genetic warfarin resistance in the rat. FASEB J. 2001;15:2542–2544.
Wajih N, Sane DC, Hutson SM, Wallin R. The inhibitory effect of calumenin on the vitamin K-dependent γ-carboxylation system: characterization of the system in normal and warfarin-resistant rats. J Biol Chem. 2004;279:25276–25283.
Esmon CT. Inflammation and thrombosis. J Thromb Haemost. 2003;1:1343–1348.
Dahlback B, Villoutreix BO. Regulation of blood coagulation by the protein C anticoagulant pathway: novel insights into structure- function relationships and molecular recognition. Arterioscler Thromb Vasc Biol. 2005;25:1311–1320.
Spek CA, Koster T, Rosendaal FR, Bertina RM, Reitsma PH. Genotypic variation in the promoter region of the protein C gene is associated with plasma protein C levels and thrombotic risk. Arterioscler Thromb Vasc Biol. 1995;15:214–218.
Aiach M, Nicaud V, Alhenc-Gelas M, et al. Complex association of protein C gene promoter polymorphism with circulating protein C levels and thrombotic risk. Arterioscler Thromb Vasc Biol. 1999;19:1573–1576.
Buil A, Soria JM, Souto JC, et al. Protein C levels are regulated by a quantitative trait locus on chromosome 16: results from the Genetic Analysis of Idiopathic Thrombophilia (GAIT) Project. Arterioscler Thromb Vasc Biol. 2004;24:1321–1325.
Mannami T, Baba S, Ogata J. Potential of carotid enlargement as a useful indicator affected by high blood pressure in a large general population of a Japanese city: the Suita Study. Stroke. 2000;31:2958- 2965.
Kokubo Y, Kamide K, Inamoto N, et al. Identification of 108 SNPs in TSC, WNK1, and WNK4 and their association with hypertension in a Japanese general population. J Hum Genet. 2004;49:507–515.
Sakata T, Okamoto A, Mannami T, Matsuo H, Miyata T. Protein C and antithrombin deficiency are important risk factors for deep vein thrombosis in Japanese. J Thromb Haemost. 2004;2:528–530.
Sakata T, Okamoto A, Mannami T, Tomoike H, Miyata T. Prevalence of protein S deficiency in the Japanese general population: the Suita Study. J Thromb Haemost. 2004;2:1012–1013.
Okuda T, Fujioka Y, Kamide K, et al. Verification of 525 coding SNPs in 179 hypertension candidate genes in the Japanese population: identification of 159 SNPs in 93 genes. J Hum Genet. 2002;47:387–394.
Antonarakis SE, and the Nomenclature Working Group. Recommendations for a nomenclature system for human gene mutations. Hum Mutat. 1998;11:1–3.
Tanaka C, Kamide K, Takiuchi S, et al. An alternative fast and convenient genotyping method for the screening of angiotensin converting enzyme gene polymorphisms. Hypertens Res. 2003;26:301–306.
Souto JC,Almasy L, Blangero J, et al. Genetic regulation of plasma levels of vitamin K-dependent proteins involved in hematostatis: results from the GAIT Project. Genetic Analysis of Idiopathic Thrombophilia. Thromb Haemost. 2001;85:88–92.
Almasy L, Soria JM, Souto JC, et al. A quantitative trait locus influencing free plasma protein S levels on human chromosome 1q: results from the Genetic Analysis of Idiopathic Thrombophilia (GAIT) Project. Arterioscler Thromb Vasc Biol. 2003;23:508–511.
Hasstedt SJ, Scott BT, Callas PW, et al. Genome scan of venous thrombosis in a pedigree with protein C deficiency. J Thromb Haemost. 2004;2:868–873.
Pudota BN, Miyagi M, Hallgren KW, et al. Identification of the vitamin K-dependent carboxylase active site: Cys-99 and Cys-450 are required for both epoxidation and carboxylation. Proc Natl Acad Sci U S A. 2000;97:13033–13038.
Mutucumarana VP, Acher F, Straight DL, Jin DY, Stafford DW. A conserved region of human vitamin K-dependent carboxylase between residues 393 and 404 is important for its interaction with the glutamate substrate. J Biol Chem. 2003;278:46488–46493.
Tie JK, Mutucumarana VP, Straight DL, Carrick KL, Pope RM, Stafford DW. Determination of disulfide bond assignment of human vitamin K-dependent γ-glutamyl carboxylase by matrix- assisted laser desorption/ionization time-of-flight mass spectrometry. J Biol Chem. 2003;278:45468–45475.
Pudota BN, Hommema EL, Hallgren KW, McNally BA, Lee S, Berkner KL. Identification of sequences within the γ-carboxylase that represent a novel contact site with vitamin K-dependent proteins and that are required for activity. J Biol Chem. 2001;276:46878–46886.
Tait RC,Walker ID, Islam SI, et al. Protein C activity in healthy volunteers: influence of age, sex, smoking and oral contraceptives. Thromb Haemost. 1993;70:281–285.
Henkens CM, Bom VJ, Van der Schaaf W, et al. Plasma levels of protein S, protein C, and factor X: effects of sex, hormonal state and age. Thromb Haemost. 1995;74:1271–1275.
Miyata T, Kimura R, Kokubo Y, Sakata T. Genetic risk factors for deep vein thrombosis among Japanese: importance of protein S K196E mutation. Int J Hematol. 2006;83:217–223.
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Kimura, R., Kokubo, Y., Miyashita, K. et al. Polymorphisms in Vitamin K—Dependent γ-Carboxylation—Related Genes Influence Interindividual Variability in Plasma Protein C and Protein s Activities in the General Population. Int J Hematol 84, 387–397 (2006). https://doi.org/10.1532/IJH97.06082
- Genetic polymorphism
- Vitamin K
- Protein C activity
- γ-Glutamyl carboxylase
- Vitamin K epoxide reductase complex subunit 1