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Thromb Haemost 2000; 84(06): 1039-1044
DOI: 10.1055/s-0037-1614168
Review Article
Schattauer GmbH

Modulation of the Binding of Matrix Gla Protein (MGP) to Bone Morphogenetic Protein-2 (BMP-2)

Reidar Wallin
1   From the Department of Internal Medicine Sections on Rheumatology, Winston-Salem, NC
,
Dean Cain
1   From the Department of Internal Medicine Sections on Rheumatology, Winston-Salem, NC
,
Susan M. Hutson
2   Department of Biochemistry, Wake Forest University School of Medicine, Winston-Salem, NC
,
David C. Sane
3   Cardiology, Winston-Salem, NC
,
Richard Loeser
4   Section on Rheumatology, Rush-Presbyterian, St. Luke’s Medical Center, Chicago, IL, USA
› Author AffiliationsThis work was supported by a grant from the National Institutes of Health (HL 60082) and partially by a grant from the Sticht Center on Aging at Wake Forest University School of Medicine. We are specifically grateful to D R. Hiskey for providing us with the Gla-peptide and to the Genetics Institute for making human recombinant BMP-2 available to us.
Further Information

Publication History

Received 10 May 2000

Accepted after revision 14 July 2000

Publication Date:
13 December 2017 (online)

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Summary

Matrix Gla protein (MGP) is an inhibitor of calcification of the arterial wall but the mechanism of inhibition has not been resolved. Since chondrogenesis has been identified in calcified arteries from MPG null mice, we hypothesized that locally produced MGP might inhibit calcification by neutralizing the known effect of bone morphogenetic proteins (BMPs) as promotors of chondrogenesis and bone formation. As the first step to test this hypothesis, we demonstrate that MGP is a binding protein for 125I-BMP-2. Optimal binding is dependent on metals which suggests that the metal binding Gla region in MGP is involved. MGP is shown to undergo a Ca++ induced conformational change despite the presence of the γ-carboxylase binding site being part of the mature protein sequence. The data propose that MGP matures earlier in the secretory pathway than other vitamin K-dependent proteins. Antibodies were used in an attempt to identify MGP in bovine serum. Conformational specific MGP antibodies were shown to also recognize the Gla region in prothrombin and factor X but did not identify MGP in serum. This finding is supported by electrophoresis data which demonstrate the absence of MGP among Ba-citrate absorbed vitamin K-dependent serum proteins. We conclude that MGP does not exist in normal bovine serum.

Keywords

Vitamin K - arterial calcification - vitamin K-dependent proteins
 

  • References

  • 1 Hauschka PV, Lian JB, Cole DE, Gundberg CM. Osteocalcin and matrix Gla protein: vitamin K-dependent proteins in bone. Physiol Rev 1989; 69: 990-1034.
    CrossrefPubMedGoogle Scholar
  • 2 Loeser R, Carlson C, Tulli H, Gray WJ, Miller L, Wallin R. Articularcartilage matrix γ-carboxyglutamic acid-containing protein: characterization and immunolocalization. Biochem J 1992; 282: 1-6.
    CrossrefPubMedGoogle Scholar
  • 3 Shanahan CM, Cary NRB, Metcalfe JC, Weissberg PL. High expression of genes for calcification-regulating proteins in human atherosclerotic plaques. J Clin Invest 1994; 93: 2393-402.
    CrossrefPubMedGoogle Scholar
  • 4 Wallin R, Cain D, Sane D. Matrix Gla protein synthesis and gammacarboxylation in the aortic vessel wall and proliferating vascular smooth muscle cells. Thromb Haemost 1999; 82: 1764-7.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 5 Price PA, Williamson MK, Haba T, Dell RB, Webster SSJ. Excessive mineralization with growth plate closure in rats on chronic warfarin treatment. Proc Natl Acad Sci USA 1982; 79: 7734-8.
    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 lamellae in rat arteries and heart valves. Arteriosclerosis, Thrombosis & Vascular Biology 1998; 18: 1400-7.
    CrossrefPubMedGoogle Scholar
  • 8 Furie B, Furie BC. Molecular and cellular biology of blood coagulation. New Engl J Med 1992; 326: 800-6.
    CrossrefPubMedGoogle Scholar
  • 9 Loeser RF, Wallin R, Sadowski J. Vitamin K and vitamin K-dependent proteins in the elderly: implications for bone and cartilage biology. In: Handbook of Nutrition in the Aged. 2nd Edition.. Watson RR. Ann Arbor: CRC Press; 1994: 263-80.
    Google Scholar
  • 10 Stanton C, Taylor R, Wallin R. Processing of prothrombin in the secretory pathway. Biochem J 1991; 277: 59-65.
    CrossrefPubMedGoogle Scholar
  • 11 Wallin R, Stanton C, Hutson SM. Intracellular maturation of the γ-carboxyglutamic acid (Gla) region in prothrombin coincides with release of the propeptide. Biochem J 1993; 291: 723-27.
    CrossrefPubMedGoogle Scholar
  • 12 Price P, Fraser JD, Metz-Virca G. Molecular cloning of matrix Gla protein: Implications for substrate recognition by the vitamin K-dependent carboxylase. Proc Natl Acad Sci USA 1987; 84: 8335-9.
    CrossrefPubMedGoogle Scholar
  • 13 Wozney JM, Rosen V, Celeste AJ, Mitsock LA, Whitters MJ, Kriz RW, Hewick RM, Wang EA. Novel regulators of bone formation: Molecular clones and activities. Science 1988; 242: 1528-34.
    CrossrefPubMedGoogle Scholar
  • 14 Riley EH, Lane JM, Urist MR, Lyons KM, Liebman JR. Bone Morphogenetic Protein-2. Clin Orthopaed Related Res 1996; 324: 39-46.
    CrossrefPubMedGoogle Scholar
  • 15 Willette RN, Gu JL, Lysko PG, Anderson KM, Minehart H, Yue T-L. BMP-2 gene expression and effects on human vascular smooth muscle cells. J Vasc Res 1999; 36: 120-5.
    CrossrefPubMedGoogle Scholar
  • 16 Price PA, Faus SA, Williamson MK. Warfarin-induced artery calcification is accelerated by growth and vitamin D. Arterioscler Throm Vasc Biol 2000; 20: 317-27.
    CrossrefPubMedGoogle Scholar
  • 17 Nijziel MR, Sniders S, Dissel P, Gijsbers BLMG, van Oerle R, Hamulyak K, Vermeer C. Serum matrix Gla protein is increased in cancer patients. Thromb Haemost Suppl August. 1999: Abstract 1636, page 520.
    PubMedGoogle Scholar
  • 18 Kuwada M, Katayama K. A high-performance liquid chromatographic method for the simultaneous determination of g-carboxyglutamic acid and glutamic acid in proteins, bone and urine. Anal Biochem 1981; 117: 259-65.
    CrossrefPubMedGoogle Scholar
  • 19 Nishimoto SK. A colorimetric assay specific for γ-carboxyglutamic acid-containing proteins: its utility in protein purification. Anal Biochem 1990; 186: 273-9.
    CrossrefPubMedGoogle Scholar
  • 20 Stenn KS, Blout ER. Mechanism of bovine prothrombin activation by an insoluble preparation of bovine factor X (thrombokinase). Biochemistry 1972; 11: 4502-15.
    CrossrefPubMedGoogle Scholar
  • 21 Bloom JW, Mann KG. Metal ion induced conformational transitions of prothrombin and prothrombin fragment 1. Biochemistry 1978; 17: 4430-8.
    CrossrefPubMedGoogle Scholar
  • 22 Hale JE, Williamson MK, Price PA. Carboxyl-terminal proteolytic processing of matrix Gla protein. J Biol Chem 1991; 266: 21145-9.
    PubMedGoogle Scholar
  • 23 Poster J, Price PA. A method for decarboxylation of γ-carboxyglutamic acid in proteins. J Biol Chem 1979; 254: 431-6.
    PubMedGoogle Scholar
  • 24 Fair DS, Bahnak BR. Human hepatoma cells secrete single chain factor X, prothrombin and antithrombin III. Blood 1984; 64: 191-204.
    PubMedGoogle Scholar
  • 25 Garnero P, Grimaux M, Seguin P, Delmas PD. Characterization of immunoreactive forms of human osteocalcin generated in vivo and in vitro. J Bone Miner Res 1994; 09: 255-64.
    PubMedGoogle Scholar
  • 26 Anderson HC. Vesicles associated with calcification in the matrix of epiphyseal cartilage. J Cell Biol 1994; 41: 59.
    PubMedGoogle Scholar
  • 27 Atkinson BL, Fantle KS, Benedict JJ, Huffer WE, Gutierrez-Hartmann A. Combination of osteoindictive bone proteins differentiates mesenchymal C3H/10T1/2 cells specifically to cartilage linage. J Cell Biol 1997; 65: 325-39.
    CrossrefPubMedGoogle Scholar
  • 28 Graff JM. Embryonic patterning: to BMP or not to BMP, that is the question. Cell 1997; 89: 171-4.
    CrossrefPubMedGoogle Scholar
  • 29 Iemura S, Yamamoto TS, Takagi C, Kobayashi H, Ueno N. Isolation and characterization of bone morphogenetic protein-binding proteins from early Xenopus embryo. J Biol Chem 1999; 274: 26843-9.
    CrossrefPubMedGoogle Scholar
  • 30 Rechler MM. Insulin-like growth factor binding proteins. Vita Horm 1993; 47: 1-114.
    PubMedGoogle Scholar
  • 31 Church WR, Meisser T, Howard PR, Amiral J, Meyer D, Mann KG. A conserved epitope on several human vitamin K-dependent proteins: localization of the antigenic site and influence of metal ions on antibody binding. J Biol Chem 1988; 263: 6259-67.
    PubMedGoogle Scholar
  • 32 Delmas PD, Stenner DD, Romberg RW, Riggs BL, Mann KG. Immunochemical studies of conformational alternations in bone gamma-carboxyglutamic acid containing proteins. Biochemistry 1984; 23: 4720-5.
    CrossrefPubMedGoogle Scholar
  • 33 Wallin R, Prydz H. Purification of bovine prothrombin by affinity chromatography. FEBS Lett 1975; 51: 191-4.
    CrossrefPubMedGoogle Scholar