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Thromb Haemost 2013; 110(06): 1135-1144
DOI: 10.1160/TH13-05-0408
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Partial deletion of the αC-domain in the Fibrinogen Perth variant is associated with thrombosis, increased clot strength and delayed fibrinolysis

Sarah K. Westbury   *
1   Bristol Heart Institute & School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK
,
Cédric Duval   *
2   Theme Thrombosis, Division of Cardiovascular and Diabetes Research, University of Leeds, Leeds, UK
,
Helen Philippou
2   Theme Thrombosis, Division of Cardiovascular and Diabetes Research, University of Leeds, Leeds, UK
,
Rebecca Brown
2   Theme Thrombosis, Division of Cardiovascular and Diabetes Research, University of Leeds, Leeds, UK
,
Kurtis R. Lee
3   Department of Haematology, University Hospitals Bristol NHS Foundation Trust, Bristol, UK
,
Sherina L. Murden
3   Department of Haematology, University Hospitals Bristol NHS Foundation Trust, Bristol, UK
,
Emma Phillips
3   Department of Haematology, University Hospitals Bristol NHS Foundation Trust, Bristol, UK
,
Christopher Reilly-Stitt
3   Department of Haematology, University Hospitals Bristol NHS Foundation Trust, Bristol, UK
,
Daniel Whalley
2   Theme Thrombosis, Division of Cardiovascular and Diabetes Research, University of Leeds, Leeds, UK
,
Robert A. Ariëns
2   Theme Thrombosis, Division of Cardiovascular and Diabetes Research, University of Leeds, Leeds, UK
,
Andrew D. Mumford
1   Bristol Heart Institute & School of Cellular and Molecular Medicine, University of Bristol, Bristol, UK
› Author Affiliations Financial support: This research was supported by the National Institute for Health Research Biomedical Research Unit in Cardiovascular Disease at the University Hospitals Bristol NHS Foundation Trust and the University of Bristol. This article/paper/report presents independent research funded by the National Institute for Health Research (NIHR). The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR or the Department of Health. RA, CD and HP receive funding from the Medical Research Council (G0901546) and the British Heart Foundation (RG/13/3/30104).
Further Information

Publication History

Received: 21 May 2013

Accepted after major revision: 07 August 2013

Publication Date:
30 November 2017 (online)

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Summary

Genetic fibrinogen (FGN) variants that are associated with bleeding or thrombosis may be informative about fibrin polymerisation, structure and fibrinolysis. We report a four generation family with thrombosis and heritable dysfibrinogenaemia segregating with a c.[1541delC];[=] variation in FGA (FGN-Perth). This deletion predicts a truncated FGN αC-domain with an unpaired terminal Cys at residue 517 of FGN-Aα. In keeping with this, SDS-PAGE of purified FGN-Perth identified a truncated FGN-Aα chain with increased co-purification of albumin, consistent with disulphide bonding to the terminal Cys of the variant FGN-Aα. Clot visco-elastic strength in whole blood containing FGN-Perth was greater than controls and tPA-mediated fibrinolysis was delayed. In FGN-Perth plasma and in purified FGN-Perth, there was markedly reduced final turbidity after thrombin-mediated clot generation. Consistent with this, FGN-Perth formed tighter, thinner fibrin fibres than controls indicating defective lateral aggregation of protofibrils. Clots generated with thrombin in FGN-Perth plasma were resistant to tPA-mediated fibrinolysis. FGN-Perth clot also displayed impaired tPA-mediated plasmin generation but incorporated α2-anti-plasmin at a similar rate to control. Impaired fibrinolysis because of defective plasmin generation potentially explains the FGN-Perth clinical phenotype. These findings highlight the importance of the FGN αC-domain in the regulation of clot formation and fibrinolysis.

Keywords

Dysfibrinogenaemia - thrombosis - fibrin polymerisation - fibrinolysis

* Joint first authors.


 

  • References

  • 1 Hill M, Dolan G. Diagnosis, clinical features and molecular assessment of the dysfibrinogenaemias. Haemophilia 2008; 14: 889-897.
    CrossrefPubMedGoogle Scholar
  • 2 Shapiro SE, Phillips E, Manning RA. et al. Clinical phenotype, laboratory features and genotype of 35 patients with heritable dysfibrinogenaemia. Br J Haematol 2012; 160: 220-227.
    CrossrefPubMedGoogle Scholar
  • 3 Cunningham MT, Brandt JT, Laposata M. et al. Laboratory diagnosis of dysfibrinogenemia. Arch Pathol Lab Med 2002; 126: 499-505.
    CrossrefPubMedGoogle Scholar
  • 4 Smith KA, Adamson PJ, Pease RJ. et al. Interactions between factor XIII and the alphaC region of fibrinogen. Blood 2011; 117: 3460-3468.
    CrossrefPubMedGoogle Scholar
  • 5 Tsurupa G, Medved L. Identification and characterisation of novel tPA- and plasminogen-binding sites within fibrin(ogen) alpha C domains. Biochemistry 2001; 40: 801-808.
    CrossrefPubMedGoogle Scholar
  • 6 Tsurupa G, Yakovlev S, McKee P. et al. Noncovalent interaction of alpha(2)-anti-plasmin with fibrin(ogen). localisation of alpha(2)-antiplasmin-binding sites. Biochemistry 2010; 49: 7643-7651.
    CrossrefPubMedGoogle Scholar
  • 7 Mosesson MW. Fibrinogen and fibrin structure and functions. J Thromb Haemost 2005; 3: 1894-1904.
    CrossrefPubMedGoogle Scholar
  • 8 Homer VM, Mullin JL, Brennan SO. et al. Novel Aalpha chain truncation (fibrinogen Perth) resulting in low expression and impaired fibrinogen polymerisation. J Thromb Haemost 2003; 1: 1245-1250.
    CrossrefPubMedGoogle Scholar
  • 9 Jayo A, Arnold E, Gonzalez-Manchon C. et al. Hypodysfibrinogenemia causing mild bleeding and thrombotic complications in a compound heterozygote of AalphaIVS4+1G>T mutation and Aalpha4841delC truncation (Aalpha(Perth)). Thromb Haemost 2009; 101: 770-772.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 10 Furlan M, Steinmann C, Jungo M. et al. A frameshift mutation in Exon V of the A alpha-chain gene leading to truncated A alpha-chains in the homozygous dysfibrinogen Milano III. J Biol Chem 1994; 269: 33129-33134.
    CrossrefPubMedGoogle Scholar
  • 11 Koopman J, Haverkate F, Grimbergen J. et al. Fibrinogen Marburg: a homozygous case of dysfibrinogenemia, lacking amino acids A alpha 461-610 (Lys 461 AAA-->stop TAA). Blood 1992; 80: 1972-1979.
    CrossrefPubMedGoogle Scholar
  • 12 Scott DJ, Prasad P, Philippou H. et al. Clot architecture is altered in abdominal aortic aneurysms and correlates with aneurysm size. Arteriosclerosis, thrombosis, and vascular biology 2011; 31: 3004-3010.
    CrossrefPubMedGoogle Scholar
  • 13 Allan P, Uitte de Willige S, Abou-Saleh RH. et al. Evidence that fibrinogen gamma' directly interferes with protofibril growth: implications for fibrin structure and clot stiffness. J Thromb Haemost 2012; 10: 1072-1080.
    CrossrefPubMedGoogle Scholar
  • 14 Schroder V, Kohler HP. Effect of factor XIII Val34Leu on alpha2-antiplasmin incorporation into fibrin. Thromb Haemost 2000; 84: 1128-1130.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 15 Bobbink IW, Tekelenburg WL, Sixma JJ. et al. Glycated proteins modulate tissue-plasminogen activator-catalysed plasminogen activation. Biochemical and biophysical research communications 1997; 240: 595-601.
    CrossrefPubMedGoogle Scholar
  • 16 Weisel JW, Nagaswami C. Computer modeling of fibrin polymerisation kinetics correlated with electron microscope and turbidity observations: clot structure and assembly are kinetically controlled. Biophys J 1992; 63: 111-128.
    CrossrefPubMedGoogle Scholar
  • 17 Wada Y, Lord ST. A correlation between thrombotic disease and a specific fibrinogen abnormality (A alpha 554 Arg-->Cys) in two unrelated kindred, Dusart and Chapel Hill III. Blood 1994; 84: 3709-3714.
    CrossrefPubMedGoogle Scholar
  • 18 Hanss M, Vergnes C, Rugeri L. et al. A new electrophoretic variant of fibrinogen associated with venous thromboembolism, fibrinogen Bordeaux Aalpha Arg439-->Cys. J Thromb Haemost 2008; 6: 1422-1424.
    CrossrefPubMedGoogle Scholar
  • 19 Brennan SO, Mosesson MW, Lowen R. et al. Dysfibrinogenemia (fibrinogen Wilmington) due to a novel Aalpha chain truncation causing decreased plasma expression and impaired fibrin polymerisation. Thromb Haemost 2006; 96: 88-89.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 20 Ridgway HJ, Brennan SO, Gibbons S. et al. Fibrinogen Lincoln: a new truncated alpha chain variant with delayed clotting. Br J Haematol 1996; 93: 177-184.
    CrossrefPubMedGoogle Scholar
  • 21 Weisel JW, Veklich Y, Gorkun O. The sequence of cleavage of fibrinopeptides from fibrinogen is important for protofibril formation and enhancement of lateral aggregation in fibrin clots. J Mol Biol 1993; 232: 285-297.
    CrossrefPubMedGoogle Scholar
  • 22 Litvinov RI, Yakovlev S, Tsurupa G. et al. Direct evidence for specific interactions of the fibrinogen alphaC domains with the central E region and with each other. Biochemistry 2007; 46: 9133-9142.
    CrossrefPubMedGoogle Scholar
  • 23 Tsurupa G, Mahid A, Veklich Y. et al. Structure, stability, and interaction of fibrin alphaC domain polymers. Biochemistry 2011; 50: 8028-8037.
    CrossrefPubMedGoogle Scholar
  • 24 Gorkun OV, Veklich YI, Medved LV. et al. Role of the alpha C domains of fibrin in clot formation. Biochemistry 1994; 33: 6986-6997.
    CrossrefPubMedGoogle Scholar
  • 25 Gorkun OV, Henschen-Edman AH, Ping LF. et al. Analysis of A alpha 251 fibrinogen: the alpha C domain has a role in polymerisation, albeit more subtle than anticipated from the analogous proteolytic fragment X. Biochemistry 1998; 37: 15434-15441.
    CrossrefPubMedGoogle Scholar
  • 26 Baradet TC, Haselgrove JC, Weisel JW. Three-dimensional reconstruction of fibrin clot networks from stereoscopic intermediate voltage electron microscope images and analysis of branching. Biophys J 1995; 68: 1551-1560.
    CrossrefPubMedGoogle Scholar
  • 27 Matsuka YV, Medved LV, Migliorini MM. et al. Factor XIIIa-catalysed cross-linking of recombinant alpha C fragments of human fibrinogen. Biochemistry 1996; 35: 5810-5816.
    CrossrefPubMedGoogle Scholar
  • 28 Carlisle CR, Sparks EA, Der Loughian C. et al. Strength and failure of fibrin fiber branchpoints. J Thromb Haemost 2010; 8: 1135-1138.
    CrossrefPubMedGoogle Scholar
  • 29 Collet JP, Woodhead JL, Soria J. et al. Fibrinogen Dusart: electron microscopy of molecules, fibers and clots, and viscoelastic properties of clots. Biophys J 1996; 70: 500-510.
    CrossrefPubMedGoogle Scholar
  • 30 Gabriel DA, Muga K, Boothroyd EM. The effect of fibrin structure on fibrinolysis. J Biol Chem 1992; 267: 24259-24263.
    CrossrefPubMedGoogle Scholar