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Thromb Haemost 2001; 86(03): 862-870
DOI: 10.1055/s-0037-1616144
Review Articles
Schattauer GmbH

Factor IX Denver, ASN 346→ASP Mutation Resulting in a Dysfunctional Protein with Defective Factor VIIIa Interaction

Jerry B. Lefkowitz
1   Department of Pathology
,
Rachelle Nuss
2   Department of Pediatrics, University of Colorado School of Medicine, Denver, CO
,
Tara Haver
1   Department of Pathology
,
Linda Jacobson
2   Department of Pediatrics, University of Colorado School of Medicine, Denver, CO
,
Arthur R. Thompson
3   Puget Sound Blood Center, and the University of Washington, Seattle, WA, USA
,
Marilyn Manco-Johnson
2   Department of Pediatrics, University of Colorado School of Medicine, Denver, CO
› Author Affiliations
Further Information

Publication History

Received 19 October 2000

Accepted after revision 25 April 2001

Publication Date:
14 December 2017 (online)

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Summary

Hemophilia B is a sex-linked recessive bleeding disorder characterized by the presence of either a decreased amount of normal factor IX (FIX) or the presence of a dysfunctional FIX. We have identified a unique mutation in a family with mild hemophilia B. DNA analysis of family members revealed a single base transition in the 8th exon of the FIX gene predicting an amino acid change of Asn 346→Asp in the catalytic domain. The FIX variant, named FIX Denver, was purified from proband plasma. Kinetic studies of factor X (FX) interactions with normal FIXa or FIXa Denver and phospholipid (PL) showed little difference in kcat, but a significant difference when factor VIIIa (FVIIIa) was included in the reaction. Using kinetic assays to infer the Kd of FIXa for FVIIIa, normal FIXa had a Kd of 0.095 nM while that of FIXa Denver was 9.85 nM. The major defect caused by this point mutation is a marked decrease in the affinity of FIXa Denver for factor VIIIa.

Keywords

Factor IX - factor VIII - factor X - hemophilia B - hemorrhage
 

  • References

  • 1 Thompson AR. Molecular biology of factor IX. In: Hemostasis and Thrombosis, Basic Principles and Clinical Practice. 4th edition. Colman RW, Hirsh J, Marder VJ, Salzman EW. editors. Philadelphia: J. B. Lipincott; in press 2000
    Search in Google Scholar
  • 2 Stenflo J, Fernlund P, Egan W, Roepstorff P. Proc Nat Acad Sci USA 1974; 71: 2730-3.
    CrossrefPubMedSearch in Google Scholar
  • 3 Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1988; 53: 505-18.
    CrossrefPubMedSearch in Google Scholar
  • 4 Bauer KA, Kass BL, ten Cate H, Hawiger JJ, Rosenberg R. Factor IX is activated in vivo by the tissue factor mechanism. Blood 1990; 76: 731-6.
    PubMedSearch in Google Scholar
  • 5 van Dieijen G, Tans G, Rosing J, Hemker HC. The role of phospholipid and factor VIIIa in the activation of bovine factor X. J Biol Chem 1981; 256: 3433-42.
    PubMedSearch in Google Scholar
  • 6 Hultin MB. Role of human factor VIII in factor X activation. J Clin Invest 1882; 69: 950-8.
    PubMedSearch in Google Scholar
  • 7 DiMichele D, Neufeld EJ. Hemophilia. A new approach to an old disease. Hematol Oncol Clin NA 1988; 12: 1315-44.
    PubMedSearch in Google Scholar
  • 8 Lozier JN, Monroe DM, Stanfield-Oakley S, Lin S-W, Smith KJ, Roberts HR, High KA. FIX New London: Substitution of proline for glutamine at position 50 causes severe hemophilia B. Blood 1990; 75: 1097-104.
    PubMedSearch in Google Scholar
  • 9 Kemball-Cook G, Tubbs JE, Dawson NJ, Barrowcliffe TW. The behaviour of different factor VIII concentrates in a chromogenic factor X-activating system. Brit J Haematol 1993; 84: 273-8.
    CrossrefPubMedSearch in Google Scholar
  • 10 Tans G, Janssen-Claessen T, van Dieijen G, Hemker HC, Rosing J. Activation of factor IX by factor XIa – A spectrophotometric assay for factor IX in human plasma. Thromb Haemost 1982; 48: 127-32.
    Thieme ConnectPubMedSearch in Google Scholar
  • 11 Weinmann AF, Murphy MEP, Thompson AR. Consequences of factor IX mutations in 26 families with haemophilia B. Brit J Haematol 1998; 100: 58-61.
    CrossrefPubMedSearch in Google Scholar
  • 12 Di Scipio RG, Hermodson MA, Yates SG, Davie EW. A comparison of human prothrombin, factor IX (Christmas factor), factor X (Stuart factor), and protein S. Biochemistry 1977; 16: 698-706.
    CrossrefPubMedSearch in Google Scholar
  • 13 Stürzebecher J, Kopetzki E, Bode W, Hopfner K-P. Dramatic enhancement of catalytic activity of coagulation factor IXa by alcohols. FEBS Letters 1997; 412: 295-300.
    CrossrefPubMedSearch in Google Scholar
  • 14 Griffith MJ, Breitkreutz L, Trapp H, Briet E, Noyes CM, Lundblad RL, Roberts HR. Characterization of the clotting activities of structurally different forms of activated factor IX. J Clin Invest 1985; 75: 4-10.
    CrossrefPubMedSearch in Google Scholar
  • 15 Hope MJ, Bally MB, Webb G, Cullis PR. Biochim Biophys Acta 1985; 812: 55-65.
    CrossrefPubMedSearch in Google Scholar
  • 16 Larson PJ, Stanfield-Oakley SA, VanDusen WJ, Kasper CK, Smith KJ, Monroe DM, High KA. Structural integrity of the γ-carboxyglutamic acid domain of human blood coagulation factor IXa is required for its binding to cofactor VIIIa. J Biol Chem 1996; 271: 3869-76.
    CrossrefPubMedSearch in Google Scholar
  • 17 Chang JY, Monroe DM, Stafford DW, Brinkhous KM, Roberts HR. Replacing the first epidermal growth factor-like domain of factor IX with that of factor VII enhances activity in vitro and in canine hemophilia B. J Clin Invest 1997; 100: 886-92.
    CrossrefPubMedSearch in Google Scholar
  • 18 High KA, Roberts HR. Factor IX. In Molecular Basis of Thrombosis and Hemostasis. K. A. High and H. R. Roberts, editors. New York: Marcel Dekker, Inc.; 215-237 1995
    Search in Google Scholar
  • 19 Zhong D, Smith KJ, Birktoft JJ, Bajaj SP. First epidermal growth factor-like domain of human blood coagulation factor IX is required for its activation by factor VIIa/tissue factor but not by factor XIa. Proc Natl Acad Sci USA 1994; 91: 3574-8.
    CrossrefPubMedSearch in Google Scholar
  • 20 Lenting PJ, Christophe OD, ter Maat H, Rees JG, Mertens K. Ca2+-binding to the first epidermal growth factor-like domain of human blood coagulation factor IX promotes enzyme activity and factor VIII light chain binding. J Biol Chem 1996; 271: 25332-7.
    CrossrefPubMedSearch in Google Scholar
  • 21 Rees DJ, Jones IM, Handford PA, Walter SJ, Esnouf MP, Smith KJ, Brownlee GG. The role of beta-hydroxyaspartate and adjacent carboxylate residues in the first EGF domain of human factor IX. EMBO J 1988; 7: 2053-61.
    PubMedSearch in Google Scholar
  • 22 Hughes PE, Morgan G, Rooney EK, Brownlee GG, Handford P. Tyrosine 69 of the first epidermal growth factor-like domain of human factor IX is essential for clotting activity. J Biol Chem 1993; 268: 17727-33.
    PubMedSearch in Google Scholar
  • 23 Mathur A, Bajaj SP. Protease and EGF1 domains of factor IXa play distinct roles in binding to factor VIIIa: Importance of helix-330 (helix-162 in chymotrypsin) of protease domain of factor IXa in its interaction with factor VIIIa. J Biol Chem 1999; 274: 18477-86.
    CrossrefPubMedSearch in Google Scholar
  • 24 McCord DM, Monroe DM, Smith KJ, Roberts HR. Characterization of the functional defect in factor IX Alabama. J Biol Chem 1990; 265: 10250-4.
    PubMedSearch in Google Scholar
  • 25 Bajaj SP, Rapaport SI, Maki SL. A monoclonal antibody to factor IX that inhibits factor VIII potentiation of factor X activation. J Biol Chem 1985; 260: 11574-80.
    PubMedSearch in Google Scholar
  • 26 Kolkman JA, Lenting PJ, Mertens K. Regions 301-303 and 333-339 in the catalytic domain of blood coagulation factor IX are factor VIII-interactive sites involved in stimulation of enzyme activity. Biochem J 1999; 339: 217-21.
    PubMedSearch in Google Scholar
  • 27 Bajaj SP. Region of factor IXa protease domain that interacts with factor VIIIa: analysis of select hemophilia B mutants. Thromb Haemost 1999; 82: 218-25.
    Thieme ConnectPubMedSearch in Google Scholar
  • 28 Mathur A, Zhong D, Sabharwal AK, Smith KJ, Bajaj SP. Interaction of factor IXa with factor VIIIa. Effects of protease domain Ca2+ binding site, proteolysis in the autolysis loop, phospholipid, and factor X. J Biol Chem 1997; 272: 23418-26.
    CrossrefPubMedSearch in Google Scholar
  • 29 Brandstetter H, Bauer M, Huber R, Lollar P, Bode W. X-ray structure of clotting factor IXa: Active site and module structure related to Xase activity and hemophilia B. Proc Natl Acad Sci USA 1995; 92: 9796-800.
    CrossrefPubMedSearch in Google Scholar
  • 30 Bottema CDK, Ketterling RP, Ii S, Yoon H-S, Phillips III JA, Sommer SA. Missense mutations and evolutionary conservation of amino acids: Evidence that many of the amino acids in factor IX function as “spacer” elements. Am J Hum Genet 1991; 49: 820-38.
    PubMedSearch in Google Scholar