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Thromb Haemost 1999; 82(06): 1639-1643
DOI: 10.1055/s-0037-1614892
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Schattauer GmbH

Fibrinogen Bastia (γ 318 Asp → Tyr) a Novel Abnormal Fibrinogen Characterized by Defective Fibrin Polymerization

Karim Chabane Lounes
1   From the Laboratoire Sainte Marie, Hôpital Hôtel-Dieu, Paris
2   Laboratoire Difema, Faculté de Medecine et de Pharmacie, Rouen
,
Claudine Soria
2   Laboratoire Difema, Faculté de Medecine et de Pharmacie, Rouen
3   INSERM U.153, Hôpital Saint Louis, Paris
,
Antoine Valognes
4   Laboratoire du Centre Hospitalier, Bastia, Corse
,
Marie France Turchini
4   Laboratoire du Centre Hospitalier, Bastia, Corse
,
Jeannette Soria
1   From the Laboratoire Sainte Marie, Hôpital Hôtel-Dieu, Paris
5   Laboratoire de Biochimie A, Hôtel-Dieu, Paris, France
,
Jaap Koopman
6   TNO-PG, Gaubius Laboratory, Leiden, The Netherlands
7   Pharming, Leiden, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 23 November 1998

Accepted after resubmission 25 May 1999

Publication Date:
10 December 2017 (online)

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Summary

A new congenital dysfibrinogen, Fibrinogen Bastia, was discovered in a 20-year-old woman with no clinical symptoms. The plasma thrombin-clotting time was severely prolonged. The functional plasma fibrinogen concentration was low (0.2 mg/ml), whereas the immunological concentration was normal (2.9 mg/ml). Purified fibrinogen Bastia displayed a markedly prolonged thrombin-clotting time related to a delayed thrombin-induced fibrin polymerization. Both the thrombin-clotting time and the fibrin polymerization were partially corrected by the addition of calcium ions. The anomaly of fibrinogen Bastia was found to be located in the γ-chain since by SDS-PAGE performed according to the method of Laemmli two γ-chains were detected, one normal and one with an apparently lower molecular weight. Furthermore, analysis of plasmin degradation products demonstrated that calcium ions only partially protect fibrinogen Bastia γ-chain against plasmin digestion, suggesting that the anomaly is located in the C-terminal part of the γ-chain. Sequence analysis of PCR-amplified genomic DNA fragments of the propositus demonstrated a single base substitution (G → T) in the exon VIII of the γ chain gene, resulting in the amino acid substitution 318 Asp (GAC) → Tyr (TAC). The PCR clones were recloned and 50% of them contained the mutation, indicating that the patient was heterozygous. These data indicate that residue Asp 318 is important for normal fibrin polymerization and the protective effect of calcium ions against plasmin degradation of the C-terminal part of the γ-chain.

Key Words

Fibrinogen - dysfibrinogenemia - plasmin degradation products - fibrin polymerization
 

  • References

  • 1 Olexa SA, Budzynski AZ. Evidence for four different polymerization sites involved in human fibrin formation. Proc Natl Acad Sci USA 1980; 77: 1374-8.
    CrossrefPubMedGoogle Scholar
  • 2 Shimizu A, Nagel GM, Doolittle RF. Photoaffinity labeling of the primary fibrin polymerization site: isolation and characterization of a labeled cyanogen bromide fragment corresponding to γ-chain residues 337-379. Proc Natl Acad Sci USA 1992; 89: 2888-92.
    CrossrefPubMedGoogle Scholar
  • 3 Spraggon G, Everse SJ, Doolittle RF. Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. Nature 1997; 389: 544-62.
    CrossrefPubMedGoogle Scholar
  • 4 Everse SJ, Spraggon G, Veerapandian L, Riley M, Doolittle RF. Crystal structure of fragment double-D from human fibrin with two different bound ligands. Biochemistry 1998; 37: 8637-42.
    CrossrefPubMedGoogle Scholar
  • 5 Hardy JJ, Carrell NA, McDonagh J. Calcium ion functions in fibrinogen conversion to fibrin. Ann NY Acad Sci 1983; 408: 279-87.
    CrossrefPubMedGoogle Scholar
  • 6 Nieuwenhuizen W, Van Ruijven-Vermeer IA, Nooijen WJ, Vermond A, Haverkate F, Hermans J. Recalculation of calcium-binding properties of human and rat fibrin(ogen) and their degradation products. Thromb Res 1981; 22: 653-7.
    CrossrefPubMedGoogle Scholar
  • 7 Dang CV, Ebert RF, Bell WR. Localization of a fibrinogen calcium binding site between g-subunit positions 311 and 336 by terbium fluorescence. J Biol Chem 1985; 260: 9713-9.
    PubMedGoogle Scholar
  • 8 Nieuwenhuizen W, Haverkate F. Calcium-binding regions in fibrinogen. Ann NY Acad Sci 1983; 408: 92-6.
    CrossrefPubMedGoogle Scholar
  • 9 Haverkate F, Timan G. Protective effect of calcium in the plasmin degradation of fibrinogen and fibrin product D. Thromb Res 1977; 10: 803-12.
    CrossrefPubMedGoogle Scholar
  • 10 Côté HCF, Lord ST, Pratt KP. γ-chain dysfibrinogenemias: molecular structure-function relationships of naturally occurring mutations in the γ-chain of human fibrinogen. Blood 1998; 92: 2195-12.
    PubMedGoogle Scholar
  • 11 Rupp C, Kuyas C, Häeberli A, Furlan M, Perret BA, Beck EA. In: Fibrinogen. Henschen A, Graeff H, Lottspeich F. eds. Berlin: Walter de Gruyter; 1982: 167-81.
    Google Scholar
  • 12 Soria J, Soria C, Samama M, Tabori S, Kehl M, Henschen A, Nieuwenhuizen W, Rimon A, Tatarski I. Fibrinogen Haifa: fibrinogen variant with absence of protective effect of calcium on plasmin degradation of gamma chains. Thromb. Haemost 1987; 57: 3103. Thromb Haemost 1987 Jun 3: 57 (3) 310-3.
    PubMedGoogle Scholar
  • 13 Koopman J, Haverkate F, Briet E, Lord ST. A congenitally abnormal fibrinogen (Vlissingen) with a 6-base deletion in the gamma-chain gene causing defective calcium binding and impaired fibrin polymerization. J Biol Chem 1991; 266: 13456-61.
    PubMedGoogle Scholar
  • 14 Yoshida N, Hirata H, Morigami Y, Imaoka S, Matsuda M, Yamazumi K, Asakura S. Characterization of an abnormal fibrinogen Osaka V with the replacement of γ-arginine 375 by glycine. The lack of high affinity calcium binding to D-domain and the lack of protective effect of calcium on fibrinolysis. J Biol Chem 1992; 267: 2753-9.
    PubMedGoogle Scholar
  • 15 Blombäck B, Blombäck M. Purification of human and bovine fibrinogen. Arkiv. Kemi 1956; 10: 415-43.
    PubMedGoogle Scholar
  • 16 Caen JP, Larrieu MJ, Samama M. L’hemostase. Methodes d’exploitation et diagnostic pratique. Expanssion scientifique; Paris: 1975
    Google Scholar
  • 17 Clauss A. Gerinnungsphysiologishe Schnellmethode zur Bestimmung des Fibrinogens. Acta Haematol (Basel) 1957; 17: 237-46.
    CrossrefPubMedGoogle Scholar
  • 18 Soria J, Soria C, Borg JY, Mirshahi M, Piguet H, Tron P, Fessard C, Caen J. Platelet aggregation occurs in congenital afibrinogenemia despite the absence of fibrinogen or its fragments in plasma and platelets, as demonstrated by immunoenzymology. Br J Haematol 1985; 60: 503-14.
    CrossrefPubMedGoogle Scholar
  • 19 Schwartz ML, Pizzo SV, Hill RL, Mc Kee PA. The effect of fibrin-stabilizing factor on the subunit structure of human fibrin. J Clin Invest 1971; 50: 1506-13.
    CrossrefPubMedGoogle Scholar
  • 20 Weber K, Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem 1969; 244: 4406-12.
    PubMedGoogle Scholar
  • 21 Laemmli UK. Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
    CrossrefPubMedGoogle Scholar
  • 22 Rixon MW, Chung BW, Davie EW. Nucleotide sequence of the gene for the γ-chain of human fibrinogen. Biochemistry 1985; 24: 2077-86.
    CrossrefPubMedGoogle Scholar
  • 23 Yoshida N, Terukina S, Okuma M, Moroi M, Aoki N, Matsuda M. Characterization of an apparently lower molecular weight γ-chain variant in fibrinogen Kyoto I. The replacement of γ-asparagine 308 by lysine which causes accelerated cleavage of fragment D1 by plasmin and the generation of a new plasmin cleavage site. J Biol Chem 1988; 263: 13848-56.
    PubMedGoogle Scholar
  • 24 Miyata T, Furukawa K, Iwanaga S, Takamatsu J, Saito H. Fibrinogen Nagoya, a replacement of glutamine-329 by arginine in the γ-chain that impairs the polymerization of fibrin monomers. J Biochem 1989; 105: 10-4.
    CrossrefPubMedGoogle Scholar
  • 25 Terukina S, Yamazumi K, Okamoto K, Yamashita H, Ito Y, Matsuda M. Fibrinogen Kyoto III: a congenital dysfibrinogen with a g aspartic acid-330 to tyrosine substitution manifesting impaired fibrin monomer polymerization. Blood 1989; 74: 2681-7.
    PubMedGoogle Scholar
  • 26 Ebert RF, Bell WR. Fibrinogen Baltimore III. Congenital dysfibrinogenemia with a shortened g-subunit. Throm Res 1988; 51: 251-8.
    PubMedGoogle Scholar
  • 27 Yoshida N, Ota K, Moroi M, Matsuda M. An apparently higher molecular weight γ-chain variant in a congenital abnormal fibrinogen Tochigi characterized by the replacement of γ arginine-275 by cysteine. Blood 1988; 71: 480-7.
    PubMedGoogle Scholar
  • 28 De Jong WW, Zweers A, Cohen LH. Influence of single amino acid substitution on electrophoretic mobility of sodium dodecyl sulfate-protein complexes. Biochem Biophys Res Commun 1978; 82: 532-9.
    CrossrefPubMedGoogle Scholar
  • 29 Noel D, Nikaido K, Ames GF. A single amino acid substitution in a histidine-transport protein drastically alters its mobility in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Biochemistry 1979; 18: 4159-65.
    CrossrefPubMedGoogle Scholar
  • 30 Varadi A, Scheraga HA. Localization of segments essential for polymerization and for calcium binding in the gamma-chain of human fibrinogen. Biochemistry 1986; 25: 519-28.
    CrossrefPubMedGoogle Scholar
  • 31 Nieuwenhuizen W, Voskuilen M, Vermond A, Haverkate F, Hermans J. A fibrinogen fragment D (D intermediate) with calcium binding but without anticlotting properties. Biochem Biophys Acta 1982; 707: 190-2.
    CrossrefPubMedGoogle Scholar
  • 32 Yee VC, Pratt KP, Côté CF, Le Trong IL, Chung DW, Davie EW, Stenkamp RE, Teller DC. Crystal structure of 30 kDa C-terminal fragment from the γ-chain of human fibrinogen. Sructure 1997; 5: 125-38.
    PubMedGoogle Scholar
  • 33 Niewenhuizen W, Gravesen M. Anticoagulant and calcium-binding properties of high molecular weight derivatives of human fibrinogen produced by plasmin (fragments X). Biochim Biophys Acta 1981; 668: 81-8.
    CrossrefPubMedGoogle Scholar
  • 34 Crabtree GR, Kant JA. Organization of the rat γ-fibrinogen gene. Alternative mRNA splice patterns produce the γA and γB (γ’) chains of fibrinogen. Cell 1982; 31: 159-66.
    PubMedGoogle Scholar
  • 35 Strong DD, Moore M, Cottrel BA, Bohonus VL, Pontes M, Evans B, Riley M, Doolittle RF. Lamprey fibrinogen γ-chain: Cloning cDNA sequencing, and general characterization. Biochemistry 1985; 24: 92-01.
    CrossrefPubMedGoogle Scholar
  • 36 Laudano AP, Doolittle RF. Influence of calcium ion on the binding of fibrin amino terminal peptides to fibrinogen. Science 1981; 212: 457-9.
    CrossrefPubMedGoogle Scholar