Download PDF
Thromb Haemost 1999; 82(06): 1764-1767
DOI: 10.1055/s-0037-1614911
Rapid Communication
Schattauer GmbH

Matrix Gla Protein Synthesis and Gamma-carboxylation in the Aortic Vessel Wall and Proliferating Vascular Smooth Muscle Cells

A Cell System which Resembles the System in Bone Cells
Reidar Wallin
1   From the Department of Internal Medicine, Sections of Rheumatology and University School of Medicine, Winston-Salem, North Carolina, USA
,
Dean Cain
1   From the Department of Internal Medicine, Sections of Rheumatology and University School of Medicine, Winston-Salem, North Carolina, USA
,
David C. Sane
2   From the Department of Internal Medicine, Sections of, Cardiology, Wake Forest University School of Medicine, Winston-Salem, North Carolina, USA
› Author AffiliationsThis work was supported by a grant from the North Carolina Biotechnology Center (Grant # 9805-ARG-0004).
Further Information

Publication History

Received 24 May 1999

Accepted after resubmission 04 August 1999

Publication Date:
10 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Matrix GLA protein (MGP) is an inhibitor of calcification in the arterial wall and its activity is dependent upon vitamin K-dependent γ-carboxylation. This modification is carried out by a warfarin sensitive enzyme system that converts specific Glu residues to γ-carboxyglutamic acid (GLA) residues. Recent studies have demonstrated that the γ-carboxylation system in the arterial wall, in contrast to that in the liver, is unable to use vitamin K as an antidote to warfarin.

By use of immunohistochemistry we demonstrate that MGP is expressed in the arterial wall and immunocytochemistry localized the MGP precursors to the endoplasmic reticulum in vascular smooth muscle cells. Resting smooth vascular muscle cells in the aortic wall and proliferating cells from explants of the aorta have all the enzymes needed for γ-carboxylation of MGP. However, when compared to the liver system, expression of the enzymes of the γ-carboxylation system in vascular smooth muscle cells is different. Of particular interest is the finding that the specific activity of the warfarin sensitive enzyme vitamin K epoxide reductase is 3-fold higher in vascular smooth muscle cells than in liver. DT-diaphorase, which catalyses the antidotal pathway for vitamin K reduction in liver, is 100-fold less active in resting vascular smooth muscle cells than in liver. Data obtained from an in vitro γ-carboxylation system suggest that the antidotal pathway catalyzed by DT-diaphorase in the vessel wall is unable to provide the carboxylase with enough reduced vitamin K to trigger γ-carboxylation of MGP. This finding provides an explanation to the inability of vitamin K to work as an antidote to warfarin intoxication of the arterial wall. Therefore the vitamin K dependent γ-carboxylation system in the arterial wall share a common feature with the system in bone cells by being unable to utilize vitamin K as an antidote.

Keywords

Matrix GLA protein - vitamin K - arterial calcification - warfarin
 

  • References

  • 1 Lian JB, Gundberg CM. Osteocalcin: Biochemical considerations and clinical applications. Clin Orthopaed and Related Research 1998; 226: 276-91.
    PubMedGoogle Scholar
  • 2 Hauschka PV, Lian JB, Cole DEC, Gundberg CM. Osteocalcin and Matrix Gla Protein: Vitamin K-Dependent Proteins in Bone. Phys Rev 1989; 69: 990-1047.
    PubMedGoogle Scholar
  • 3 Urist MR, Huo YK, Brownell AG, Hohl WM, Buyske J, Lietze A, Tempst P, Hunkapiller M, LeLange RJ. Purification of bovine bone morphogenetic protein by hydroxyapatite chromatography. Proc Nat Acad Sci USA 1984; 81: 371-5.
    CrossrefPubMedGoogle Scholar
  • 4 Fraser JD, Price PA. Lung, heart and kidney express high levels of mRNA for the vitamin K-dependent matrix Gla protein: implications for the possible function of matrix Gla protein and for the tissue distribution of the γ-carboxylase. J Biol Chem 1988; 263: 11033-6.
    PubMedGoogle Scholar
  • 5 Desbois DP, Boyce B, Pinero G, Dunstan C, Smith E, Bonadio J, Goldstein S, Gundberg C, Bradley A, Karsenty G. Increased bone formation in osteocalcin-deficient mice. Nature 1996; 382: 448-52.
    CrossrefPubMedGoogle Scholar
  • 6 Luo G, Ducy P, McKee MD, Pinero GJ, Loyer E, Behringer RR, Karsinsky G. Spontaneous calcification of arteries and cartilage in mice lacking matrix Gla protein. Nature 1997; 386: 78-81.
    CrossrefPubMedGoogle Scholar
  • 7 Price PA, Faus SA, Williamson MK. Warfarin causes rapid calcification of the elastic lamella in rat arteries and heart valves. Arterioscler Thromb Vasc Biol 1998; 18: 1400-7.
    CrossrefPubMedGoogle Scholar
  • 8 Wallin R, Suttie JW. Vitamin K-dependent carboxylation and vitamin K epoxidation: evidence that the warfarin-insensitive microsomal NAD(P)H dehydrogenase reduces vitamin K in these reactions. Biochem J 1980; 194: 983-8.
    PubMedGoogle Scholar
  • 9 Wallin R, Martin LF. Warfarin poisoning and vitamin K antagonism in rat and human livers: design of a system in vitro that mimics the system in vivo. Biochem J 1987; 241: 389-96.
    CrossrefPubMedGoogle Scholar
  • 10 Cain D, Hutson SM, Wallin R. Warfarin resistance is associated with a protein component of the vitamin K 2,3-epoxide reductase enzyme complex in rat liver. Thromb Haemost 1998; 80: 128-33.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 11 Wallin R, Rannels SR. Identification of vitamin K-dependent carboxylase activity in lung type II cells but not in lung macrophages. Biochem J 1988; 250: 557-63.
    CrossrefPubMedGoogle Scholar
  • 12 Freeman EJ, Chisolm GM, Ferrario CM, Tallant EA. Angiotensin-(1-7) inhibits vascular smooth muscle cell growth. Hypertension 1996; 28: 104-8.
    CrossrefPubMedGoogle Scholar
  • 13 Wallin R, Guenthner TM. Purification of warfarin-sensitive vitamin K epoxide reductase. Meth Enzymol 1997; 283 Part L 395-408.
    PubMedGoogle Scholar
  • 14 Wallin R, Martin LF. Vitamin K-dependent carboxylation and vitamin K metabolism in liver. J Clin Invest 1985; 76: 1879-84.
    CrossrefPubMedGoogle Scholar
  • 15 Wallin R, Martin LF. Early processing of prothrombin and factor X by the vitamin K-dependent carboxylase. J Biol Chem 1988; 263: 9994-10001.
    PubMedGoogle Scholar
  • 16 Ernster L. DT-diaphorase: Meth Enzymol. 1967; 10: 309-17.
    PubMedGoogle Scholar
  • 17 Price PA, Williamson MK. Primary structure of bone matrix Gla protein, a new vitamin K-dependent bone protein. J Biol Chem 1985; 260: 14917-75.
    PubMedGoogle Scholar
  • 18 Loeser R, Carlson C, Tulli H, Jerome G, Miller L, Wallin R. Articular cartilage matrix Gla protein: characterization and immunolocalization. Biochem J 1991; 287: 1-6.
    PubMedGoogle Scholar
  • 19 Pan LC, Williamson MK, Price PA. Sequence of the precursor to rat bone γ-carboxyglutamic acid protein that accumulates in warfarin–treated osteosarcoma cells. J Biol Chem 1985; 260: 13398-401.
    PubMedGoogle Scholar
  • 20 Suttie JW, Lehrman SR, Geweke LO, Hageman JM, Rich DH. Vitamin K-dependent carboxylase: requirements for carboxylation of soluble peptide substrates and substrate specificity. Biochem Biophys Res Commun 1979; 86: 500-7.
    CrossrefPubMedGoogle Scholar
  • 21 Shanahan CM. Proudfoot Farzaneh-far A, Weissberg PL. The role of GLA proteins in vascular calcification. Critical Review in Eukaryotic Gene Expression 1998; 8: 357-75.
    PubMedGoogle Scholar
  • 22 Cancela L, Hsieh C-L, Francke U, Price PA. Molecular structure, chromosome assignment, and promotor organization of the human matrix Gla protein gene. J Biol Chem 1990; 265: 15040-8.
    PubMedGoogle Scholar
  • 23 Van Haarlem JM, Ulrich MMW, Hemker C, Soute BAM, Vermeer C. Isolation and partial characterization of a vitamin K-dependent carboxylase from bovine aorta. Biochem J 1987; 245: 251-5.
    CrossrefPubMedGoogle Scholar
  • 24 Shanahan CM, Cary NRB, Weissberg PL, Edmonds ME. Medial localization of calcification-regulating proteins in association with Monckeberg’s sclerosis. Circulation 1997; 96: 3722.
    PubMedGoogle Scholar
  • 25 Cain D, Hutson SM, Wallin R. Assembly of the warfarin sensitive vitamin K-2,3-epoxidase reductase enzyme complex in the endoplasmic reticulum-membrane. J Biol Chem 1997; 272: 29068-75.
    CrossrefPubMedGoogle Scholar
  • 26 Canfield L, Davy LA, Thomas GL. Anti-oxidant/pro-oxidant reactions of vitamin K. Biochem Biophys Res Commun 1985; 128: 211-9.
    CrossrefPubMedGoogle Scholar
  • 27 Vervoort LM, Ronden JE, Thijssen HH. The potent antioxidant activity of the vitamin K cycle in microsomal lipid peroxidation. Biochem Pharmacol 1997; 54: 871-6.
    CrossrefPubMedGoogle Scholar
  • 28 Ronden JE, Drittij-Reinders M-J, Vermeer C, Thijssen HHW. Intestinal flora is not an intermediate in the phylloquinone-menaquinone-4 conversion in the rat. Biochem Biophys Acta 1998; 1379: 69-75.
    CrossrefPubMedGoogle Scholar
  • 29 Price PA, Kaneda Y. Vitamin K counteracts the effect of warfarin in liver but not in bone. Thromb Res 1987; 46: 121-31.
    CrossrefPubMedGoogle Scholar