Download PDF
Thromb Haemost 1998; 79(06): 1136-1143
DOI: 10.1055/s-0037-1615030
Rapid Communication
Schattauer GmbH

Characterization of a Factor VII Molecule Carrying a Mutation in the Second Epidermal Growth Factor-like Domain

Anita Kavlie
1   From the Biotechnology Centre of Oslo, University of Oslo
,
Lars Örning
2   Nycomed Pharma A/S, Oslo, Norway
,
Anne Grindflek
1   From the Biotechnology Centre of Oslo, University of Oslo
,
Helge Stormorken
3   Research Institute for Internal Medicine, University of Oslo, Oslo, Norway
,
Hans Prydz
1   From the Biotechnology Centre of Oslo, University of Oslo
› Author Affiliations
Further Information

Publication History

Received 18 July 1997

Accepted after resubmission 04 February 1998

Publication Date:
07 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

A missense mutation at codon 100 in the second epidermal growth factor-like domain, resulting in Gln100→Arg, was detected in 19 out of 21 available severely factor VII (FVII) deficient patients in Norway. Seventeen patients were homozygous, and the two remaining were compound heterozygotes. In the homozygous patients, FVII antigen was measured to 10-28%, and activity to 0.6-6.5% of that in normal pooled plasma. Recombinant FVII containing the mutation was expressed transiently in CHO cells to a mean antigen level of 57% of the wild type FVII protein, and with a specific activity of 6% of wild type. The mutant protein had a 14-fold reduction in affinity for tissue factor (TF), whereas binding of FX seemed unaffected. In line with the experimental data, molecular modelling of the mutation based on the coordinates of the tissue factor/FVIIa complex showed that substituting arginine for glutamine disrupts the interface between the catalytic and second epidermal growth factor-like domains.

 

  • References

  • 1 Alexander B, Goldstein R, Landwehr G. The prothrombin conversion accelerator of serum (SPCA): its partial purification and its properties compared with serum Ac-globulin. J Clin Invest 1950; 29: 881.
    CrossrefPubMedGoogle Scholar
  • 2 Owren PA. Proconvertin, the new clotting factor. Scand J Clin Lab Invest 1951; 3: 168a.
    CrossrefPubMedGoogle Scholar
  • 3 Koller F, Loeliger A, Duckert F. Experiments on a new clotting factor (factor VII). Acta Haematol 1951; 6: 1.
    CrossrefPubMedGoogle Scholar
  • 4 Gladhaug Å, Prydz H. Purification of the coagulation factors VII and X from human serum. Some properties of Factor VII Bioch Biophys Acta 1970; 215: 105.
    PubMedGoogle Scholar
  • 5 Hagen F, Gray CL, O’Hara P, Grant FJ, Saari GC, Woodbury RG, Hart CE, Insley M, Kisiel W, Kurachi K, Davie E. Characterization of a cDNA coding for human factor VII. Proc Natl Acad Sci USA 1986; 83: 2412.
    CrossrefPubMedGoogle Scholar
  • 6 O’Hara P, Grant FJ, Haldemann BJ, Gray CL, Insley MJ, Hagen FS, Murray MJ. Nucleotide sequence of the gene coding for human factor VII, a vitamin K-dependent protein participating in blood coagulation. Proc Natl Acad Sci USA 1987; 84: 5158.
    CrossrefPubMedGoogle Scholar
  • 7 Dickinson CD, Kelly CR, Ruf W. Identification of surface residues mediating tissue factor binding and catalytic function of the serine protease factor VIIa. Proc Natl Acad Sci USA 1996; 93: 14379.
    CrossrefPubMedGoogle Scholar
  • 8 Kazama Y, Patsuzyn A, Wildgoose P, Hamamoto T, Kisiel W. Isolation and characterization of proteolytic fragments of human factor VIIa which inhibit the tissue factor enhanced amidolytic activity of Factor VIIa. J Biol Chem 1993; 268: 16231.
    PubMedGoogle Scholar
  • 9 Toomey JR, Smith KJ, Stafford DW. Localization of the human tissue factor recognition determinant of human factor VIIa. J Biol Chem 1991; 266: 19198.
    PubMedGoogle Scholar
  • 10 Clarke BJ, Ofosu FA, Sridhara S, Bona RD, Rickles FR. Blajchman MA The first epidermal growth factor domain of human coagulation factor VII is essential for binding with tissue factor. FEBS Letters 1992; 298: 206.
    CrossrefPubMedGoogle Scholar
  • 11 Kumar A, Blumenthal DK, Fair DS. Identification of molecular sites on factor VII which mediate its assembly and function in the extrinsic pathway activation complex. J Biol Chem 1991; 266: 915.
    PubMedGoogle Scholar
  • 12 Kumar A, Fair D. Specific molecular interaction sites on factor VII involved in factor X activation. Eur J Biochem 1993; 217: 509.
    CrossrefPubMedGoogle Scholar
  • 13 Banner DW, D’Arcy A, Chène C, Winkler FK, Guha A, Konigsberg WH, Nemerson Y, Kirchhofer D. The crystal structure of the complex of blood coagulation factor VIIa with soluble tissue factor. Nature 1996; 380: 41-6.
    CrossrefPubMedGoogle Scholar
  • 14 Chen Q, Clarke BJ, Blajchman MA, Ofusu FA. Factor VII Hamilton: a novel type 2 mutation located at residue 57 in the first EGF domain of human factor VII. Thromb Haemost 1993; 69: 1262.
    PubMedGoogle Scholar
  • 15 Chaing S, Clarke B, Sridhara S, Chu K, Friedman P, Van Dusen W, Roberts HR, Blajchman M, Monroe DM, High KA. Severe factor VII deficiency caused by mutations abolishing the cleavage site for activation and altering binding to tissue factor. Blood 1994; 83: 3524.
    PubMedGoogle Scholar
  • 16 Takamiya O, Kemball-Cook G, Martin DM, Cooper DN, von Felten A, Meili E, Hann I, Prangnell DR, Lumley H, Tuddenham EGD, McVey JH. Detection of missense mutations by single-strand conformational polymorphism (SSCP) analysis in five dysfunctional variants of coagulation factor VII. Hum Mol Genet 1993; 2: 1355.
    CrossrefPubMedGoogle Scholar
  • 17 McVey JH, Takamiya O, Tamagnini G, Valente V, Fidalgo T, Layton M, Tuddenham EGD. Exclusion of the first EGF 1 domain of factor VII by a splice site mutation causes lethal factor VII deficiency. Thromb Haemost 1995; 73: 2061.
    PubMedGoogle Scholar
  • 18 Bernardi F, Patracchini P, Gemmati D, Ferrati M, Arcieri P, Papacchini M, Redaelli R, Baudo F, Mariani G, Marchetti G. Molecular analysis of factor VII deficiency in Italy: a frequent mutation (FVII Lazio) in a repeated intronic region. Hum Genet 1993; 92: 446.
    CrossrefPubMedGoogle Scholar
  • 19 Kavlie A, Wright MS, Stormorken H, Prydz H. Mutations in the Factor VII gene of Norwegian FVII deficient patients. Thromb Haemost 1993; 69: 1262.
    PubMedGoogle Scholar
  • 20 Kuppuswamy MN, Sabharwal AK, Birktoft JJ, Bajaj SP. Molecular characterization of human factor VII Kansas (GK704): substitution of Gln100 by Arg in one allele and of Arg304 by Gln possibly in the other allele. Thromb Haemost 1993; 69: 1292.
    PubMedGoogle Scholar
  • 21 Marchetti G, Patracchini P, Papacchini M, Ferrati M, Bernardi F. A polymorphism in the 5’ region of coagulation factor VII gene (F7) caused by an inserted decanucleotide. Hum Genet 1992; 90: 575.
    PubMedGoogle Scholar
  • 22 Bernardi F, Arcieri P, Bertina RM, Chiarotti F, Corral J, Pinotti M, Prydz H, Samama M, Sandset PM, Strom R, Vicente Garcia V, Mariani G. Contribution of Factor VII genotype to activated FVII levels. Differences in Geno-type Frequencies between Northern and Southern European populations. Arteriosclerosis, Thrombosis and Vascular Biology 1997; 17: 2548-53.
    CrossrefPubMedGoogle Scholar
  • 23 Marchetti G, Patracchini P, Gematti D, DeRosa V, Pinotti M, Rodorigo G, Casonato A, Girolami A, Bernardi F. Detection of two missense mutations and characterization of a repeat polymorphism in the factor VII gene (F7). Hum Genet 1992; 130.
    PubMedGoogle Scholar
  • 24 Marchetti G, Ferrati M, Patracchini P, Redaelli R, Bernardi F. A missense mutation (178Cys→Tyr) and two neutral dimorphisms (115His and 333Ser) in the human coagulation factor VII gene. Hum Mol Genet 1993; 2: 1055.
    CrossrefPubMedGoogle Scholar
  • 25 Green F, Kelleher C, Wilkes H, Temple A, Meade T, Humphries SE. A common genetic polymorphism associated with lower coagulation factor VII levels in healthy individuals. Arterioscler Thromb 1991; 11: 540.
    CrossrefPubMedGoogle Scholar
  • 26 Humphries SE, Lane A, Dawson S, Green F. The study of gene-environment interactions that influence thrombosis and fibrinolysis. Genetic variation at the loci for factor VII and plasminogen activator inhibitor-1. Arch Pathol Lab Med 1992; 12: 1322.
    PubMedGoogle Scholar
  • 27 Barstad RM, Stormorken H, Örning L, Stephens RW, Petersen LB, Kierulf P, Sakariassen K. Reduced thrombus formation in native blood of homozygous factor VII-deficient patients at high arterial wall shear rate. Blood 1994; 84: 3371-7.
    PubMedGoogle Scholar
  • 28 Hultman T, Bergh S, Moks T, Uhlen M. Bidirectional solid-phase sequencing of in vitro-amplified plasmid DNA. Biotechniques 1991; 10: 84.
    PubMedGoogle Scholar
  • 29 Ansorge W, Sproat B, Stegemann J, Schwager C, and Zenke M. Automated DNA sequencing: ultrasensitive detection of fluorescent bands during electrophoresis. Nucleic Acids Res 1987; 15: 4593.
    CrossrefPubMedGoogle Scholar
  • 30 Ansorge W, Voss H, Wiemann S, Schwager C, Sproat B, Zimmermann J, Stegemann J, Erfle H, Hewitt NH, Rupp T. High-throughput automated DNA sequencing facility with fluorescent labels at European Molecular Biology Laboratory. Electrophoresis 1992; 13: 616.
    CrossrefPubMedGoogle Scholar
  • 31 Orita M, Iwahan H, Kanazawa H, Hayashi K, Sekiya T. Detection of polymorphisms of human DNA by gel electrophoresis as single strand conformation polymorphisms. Proc Natl Acad Sci USA 1989; 86a: 2766.
    PubMedGoogle Scholar
  • 32 Orita M, Suzuki Y, Sekiya T, Hayashi K. Rapid and sensitive detection of point mutations and DNA polymorphisms using the polymerase chain reaction. Genomics 1989; 5b: 874.
    PubMedGoogle Scholar
  • 33 Hjort PE. Intermediate reactions in the coagulation of blood with tissue thromboplastin. Scand J Clin Lab Invest 1957; 9: 183.
    PubMedGoogle Scholar
  • 34 O’Brien DP, Gale K, Anderson JS, McVey JH, Meade T, Miller G, Tuddenham EGD. Purification and characterization of Factor VII 304-Gln: A variant molecule with reduced activity isolated from a clinically unaffected male. Blood 1991; 78: 132-40.
    PubMedGoogle Scholar
  • 35 Rao LV, Williams T, Rapaport SI. Studies of the activation of factor VII bound to tissue factor. Blood 1996; 87: 3738-48.
    PubMedGoogle Scholar
  • 36 Fang CH, Lin TC, Guha A, Nemerson Y, Konigsberg WH. Activation of factor X by factor VIIa complexed with human-mouse tissue factor chimeras requires human exon 3. Thromb Haemost 1996; 76: 361-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 37 Kelley RF, Costas KE, O’Connell MP, Lazarus RA. Analysis of the factor VIIa binding site on human tissue factor: Effects of tissue factor mutations on the kinetics and thermodynamics of binding. Biochemistry 1995; 34: 10383-92.
    CrossrefPubMedGoogle Scholar