Download PDF
Thromb Haemost 1999; 81(02): 268-274
DOI: 10.1055/s-0037-1614456
Review Articles
Schattauer GmbH

Fibrinogen St. Gallen I (γ 292 Gly → Val): Evidence for Structural Alterations Causing Defective Polymerization and Fibrinogenolysis

Bettina Stucki
1   From the Central Hematology Laboratory, Inselspital, University Hospital, Bern and, Inselspital, University Hospital, Bern, Switzerland
,
Peter Schmutz
2   Thrombosis Laboratory, Department of Internal Medicine, Inselspital, University Hospital, Bern, Switzerland
,
Luzius Schmid
3   Institute of Clinical Chemistry and Hematology, Kantonsspital, St. Gallen, Switzerland
,
André Haeberli
2   Thrombosis Laboratory, Department of Internal Medicine, Inselspital, University Hospital, Bern, Switzerland
,
Bernhard Lämmle
1   From the Central Hematology Laboratory, Inselspital, University Hospital, Bern and, Inselspital, University Hospital, Bern, Switzerland
,
Miha Furlan
1   From the Central Hematology Laboratory, Inselspital, University Hospital, Bern and, Inselspital, University Hospital, Bern, Switzerland
› Author AffiliationsThis study was supported by Grants No. 32-47033.96 and 31-33807.92 from the Swiss National Science Foundation.
Further Information

Publication History

Received10 September 1998

Accepted after revision29 October 1998

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

Fibrinogen St. Gallen I was detected in an asymptomatic Swiss woman. Routine coagulation tests revealed a prolonged thrombin and reptilase time. Functionally measured fibrinogen levels were considerably lower than those determined immunologically. Polymerization of fibrin monomers derived from purified fibrinogen was delayed in the presence of either calcium or EDTA. Normal fibrinopeptide A and B release by thrombin was established. An abnormal degradation of fibrinogen St. Gallen I by plasmin was observed. Fragment D1 of normal fibrinogen was fully protected against further proteolysis in the presence of 10 mM calcium, whereas fibrinogen St. Gallen I was partially further degraded to fragments D2 and D3. In the presence of 10 mM EDTA, the conversion of variant fragment D1 to D2 was accelerated whereas the degradation of fragment D2 to D3 was delayed in comparison to degradation of fragments D1 and D2 of normal fibrinogen. Three high-affinity calcium binding sites were found in both normal and variant fibrinogen. Mutation screening with SSCP analysis suggested a mutation in exon VIII of the → Val) as previously described in fibrinogen Baltimore I (4, 5). Dysfibrinogen St. Gallen I is characterized by delayed polymerization and abnormal fibrinogenolysis compared to normal fibrinogen. Computer modeling of the variant fibrinogen revealed structural changes that may explain its abnormal functional behaviour.γ-chain gene. Cycle sequencing of this gene portion revealed a single base substitution from G to T of the base 7527, leading to replacement of γ 292 glycine by valine. The same mutation has already been described for the fibrinogen variant Baltimore I. Molecular modeling was performed of a part of the γ-chain containing the mutation site, based on recently published X-ray crystal structures of human fibrinogen fragment D and of a 30 kD C-terminal part of the γ-chain. Significant structural alterations due to the substitution of glycine by valine at γ 292 were observed, e.g. spreading of the protein backbone, probably leading to a modified accessibility of the plasmic cleavage sites in the γ-chain at 356 Lys and 302 Lys. A shift of γ 297 Asp that is involved in interactions of fragment D with the Gly-Pro-Arg-Pro-peptide was noted by molecular modeling. The latter observation is compatible with delayed polymerization of fibrin monomers.

 

  • References

  • 1 Ebert RF. Index of variant human fibrinogens. Boca Raton, Ann Arbor, Boston: CRC Press; 1991
    Google Scholar
  • 2 Yee VC, Pratt KP, Côté HCF, Le Trong I, Chung DW, Davie EW, Stenkamp RE, Teller DC. Crystal structure of a 30 kDa C-terminal fragment from the γ chain of human fibrinogen. Structure 1997; 5: 125-38.
    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: 455-62.
    CrossrefPubMedGoogle Scholar
  • 4 Beck EA, Charache P, Jackson DP. A new inherited coagulation disorder caused by an abnormal fibrinogen (“fibrinogen Baltimore”). Nature 1965; 208: 143-5.
    CrossrefPubMedGoogle Scholar
  • 5 Bantia S, Mane SM, Bell WR, Dang CV. Fibrinogen Baltimore I: Polymerization defect associated with a γ292Gly → Val (GGC → GTC) mutation. Blood 1990; 76: 2279-83.
    PubMedGoogle Scholar
  • 6 Clauss A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta Haematol 1957; 17: 237-46.
    CrossrefPubMedGoogle Scholar
  • 7 Schulz FH. Eine einfache Bewertungsmethode von Leberparenchymschäden (volumetrische Fibrinbestimmung). Acta Hepatol 1955; 3: 306-10.
    PubMedGoogle Scholar
  • 8 Laurell CB. Quantitative estimation of proteins by electrophoresis in agarose gel containing antibodies. Anal Biochem 1966; 15: 45-52.
    CrossrefPubMedGoogle Scholar
  • 9 Furlan M, Rupp C, Beck EA, Svendsen L. Effect of calcium and synthetic peptides on fibrin polymerization. Thromb Haemost 1982; 47: 118-21.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Bögli C, Hofer A, Baudo F, Redaelli R, Furlan M. Fibrinogen Milano IV, another case of congenital dysfibrinogenemia with an abnormal fibrinopeptide A release (Aα 16 Arg → His). Haemostasis 1992; 22: 7-11.
    PubMedGoogle Scholar
  • 11 Kehl M, Lottspeich F, Henschen A. Analysis of human fibrinopeptides by high-performance liquid chromatography. Hoppe Seylers Z Physiol Chem 1981; 362: 1661-4.
    PubMedGoogle Scholar
  • 12 Furlan M, Stucki B, Steinmann C, Jungo M, Lämmle B. Normal binding of calcium to five fibrinogen variants with mutations in the carboxy terminal part of the γ-chain. Thromb Haemost 1996; 76: 377-83.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 13 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-5.
    CrossrefPubMedGoogle Scholar
  • 14 Scatchard G. The attractions of proteins for small molecules and ions. Ann NY Acad Sci 1949; 51: 660-72.
    CrossrefPubMedGoogle Scholar
  • 15 Steinmann C, Bögli C, Jungo M, Lämmle B, Heinemann G, Wermuth B, Redaelli R, Baudo F, Furlan M. Fibrinogen Milano V: a congenital dysfibrinogenaemia with a γ 275 Arg → Cys substitution. Blood Coag Fibrinol 1994; 5: 463-71.
    PubMedGoogle Scholar
  • 16 Pratt KP, Côté HCF, Chung DW, Stenkamp RE, Davie EW. The primary fibrin polymerization pocket: Three-dimensional structure of a 30-kDa C-terminal γ chain fragment complexed with the peptide Gly-Pro-Arg-Pro. Proc Natl Acad Sci USA 1997; 94: 7176-81.
    CrossrefPubMedGoogle Scholar
  • 17 Henschen A, McDonagh J. Fibrinogen, fibrin and factor XIII. In: Zwaal and Hemker (eds). Blood coagulation. Amsterdam: Elsevier 1986; 171-241.
    PubMedGoogle Scholar
  • 18 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: 310-3.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 19 Koopman J, Haverkate F, Briët E, Lord ST. A congenitally abnormal fibrinogen (Vlissingen) with a 6-base deletion in the γ-chain gene, causing defective calcium binding and impaired fibrin polymerization. J Biol Chem 1991; 266: 13456-61.
    PubMedGoogle Scholar
  • 20 Rupp C, Kuyas C, Häberli A, Furlan M, Perret BA, Beck EA. Fibrinogen Bern I: a hereditary fibrinogen variant with defective conformational stabilization by calcium ions. In: Henschen, Graeff and Lottspeich (eds). Fibrinogen – recent biochemical and medical aspects. Berlin, New York: Walter de Gruyter 1982; 167-81.
    PubMedGoogle Scholar
  • 21 Steinmann C, Reber P, Jungo M, Lämmle B, Heinemann G, Wermuth B, Furlan M. Fibrinogen Bern I: substitution γ 337 Asn → Lys is responsible for defective fibrin monomer polymerization. Blood 1993; 82: 2104-8.
    PubMedGoogle Scholar
  • 22 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. J Biol Chem 1992; 267: 2753-9.
    PubMedGoogle 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. J Biol Chem 1988; 263: 13848-56.
    PubMedGoogle Scholar
  • 24 Yoshida N, Imaoka S, Hirata H, Matsuda M, Asakura S. Heterozygous abnormal fibrinogen Osaka III with the replacement of γ arginine-275 by histidine has an apparently higher molecular weight γ-chain variant. Thromb Haemost 1992; 68: 534-8.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 25 Everse SJ, Spraggon G, Doolittle RF. A three-dimensional consideration of variant human fibrinogens. Thromb Haemost 1998; 80: 1-9.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 26 Yamazumi K, Doolittle RF. Photoaffinity labeling of the primary fibrin polymerization site: localization of the label to γ-chain Tyr-363. Proc Natl Acad Sci USA 1992; 89: 2893-6.
    CrossrefPubMedGoogle Scholar
  • 27 Shimizu A, Nagel GM, Doolittle RF. Photoaffinity labeling of the primary fibrin-polymerization site: isolation of a cyanogen bromide fragment corresponding to γchain residues 337-379. Proc Natl Acad Sci USA 1992; 89: 2888-92.
    CrossrefPubMedGoogle Scholar
  • 28 Mosesson MW. Fibrinogen and fibrin polymerization: appraisal of the binding events that accompany fibrin generation and fibrin clot assembly. Blood Coag Fibrinol 1997; 8: 257-67.
    CrossrefPubMedGoogle Scholar