Download PDF
Thromb Haemost 2003; 90(06): 1021-1028
DOI: 10.1160/TH03-06-0418
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Fibrinogen Bβ polymorphisms do not directly contribute to an altered in vitro clot structure in humans

Ghassan J. Maghzal
1   Molecular Pathology, Canterbury Health Laboratories, Christchurch, New Zealand
,
Stephen O. Brennan
1   Molecular Pathology, Canterbury Health Laboratories, Christchurch, New Zealand
,
Peter M. George
1   Molecular Pathology, Canterbury Health Laboratories, Christchurch, New Zealand
› Author AffiliationsFinancial support: This research was supported by a grant from the National Heart Foundation of New Zealand.
Further Information

Publication History

Received 30 June 2003

Accepted after resubmission 07 September 2003

Publication Date:
05 December 2017 (online)

Also available at
    Permissions and Reprints

Summary

Fibrinogen Bβ polymorphisms, such as the -455 G/A and the Arg448Lys amino acid substitution, have been shown to increase the risk of atherothrombotic disease. Although these polymorphisms are related to fibrinogen concentrations, their effect on fibrin clot structure has not been extensively studied. We examined the frequency of the fibrinogen Bβ -455 G/A polymorphism in a group of myocardial infarction (MI) patients. There was no association between this polymorphism and MI. However, we found that patients homozygous for the rare -455 A allele had a higher average age at first MI. A similar result was found for individuals homozygous for the Bβ 448 Lys allele who also had a higher age at first MI.We subsequently studied the clotting properties of purified Arg448 and Lys448 fibrinogens in vitro and found that these fibrinogens did not significantly differ in their polymerisation, fibrinolysis kinetics or in their clot permeation properties. Mass spectrometry analysis of endoproteinase Asp-N digests of Bβ chains revealed that the Lys448 and the Arg448 chains were expressed in approximately equal proportions in a heterozygote Arg448Lys individual. Our results demonstrate that the fibrinogen Bβ –455 G/A polymorphism is not associated with myocardial infarction and furthermore the closely linked Bβ Arg448Lys protein coding variation does not have an influence on the function nor the structure of the protein in a purified system.

Keywords

Atherosclerosis - fibrinogen / fibrin - polymorphisms - clot structure - expression
 

  • References

  • 1 Weisel JW, Stauffacher CV, Bullitt E. et al. A model for fibrinogen: domains and sequence. Science 1985; 230: 1388-91.
    CrossrefPubMedGoogle Scholar
  • 2 Ariens RA, Philippou H, Nagaswami C. et al. The factor XIII V34L polymorphism accelerates thrombin activation of factor XIII and affects cross-linked fibrin structure. Blood 2000; 96: 988-95.
    PubMedGoogle Scholar
  • 3 Woodhead JL, Nagaswami C, Matsuda M. et al. The ultrastructure of fibrinogen Caracas II molecules, fibers, and clots. J Biol Chem 1996; 271: 4946-53.
    CrossrefPubMedGoogle Scholar
  • 4 Wolberg AS, Monroe DM, Roberts HR. et al. Elevated prothrombin results in clots with an altered fiber structure: a possible mechanism of the increased thrombotic risk. Blood 2003; 101: 3008-13.
    CrossrefPubMedGoogle Scholar
  • 5 Blomback B, Carlsson K, Hessel B. et al. Native fibrin gel networks observed by 3D microscopy, permeation and turbidity. Biochim Biophys Acta 1989; 997: 96-110.
    CrossrefPubMedGoogle Scholar
  • 6 Thomas AE, Green FR, Kelleher CH. et al. Variation in the promoter region of the beta fibrinogen gene is associated with plasma fibrinogen levels in smokers and nonsmokers. Thromb Haemost 1991; 65: 487-90.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 7 Montgomery HE, Clarkson P, Nwose OM. et al. The acute rise in plasma fibrinogen concentration with exercise is influenced by the G-453-A polymorphism of the beta-fibrinogen gene. Arterioscler Thromb Vasc Biol 1996; 16: 386-91.
    CrossrefPubMedGoogle Scholar
  • 8 Behague I, Poirier O, Nicaud V. et al. Beta fibrinogen gene polymorphisms are associated with plasma fibrinogen and coronary artery disease in patients with myocardial infarction. The ECTIM Study. Etude Cas-Temoins sur l’Infarctus du Myocarde. Circulation 1996; 93: 440-9.
    CrossrefPubMedGoogle Scholar
  • 9 Thomas AE, Green FR, Humphries SE. Association of genetic variation at the beta-fibrinogen gene locus and plasma fibrinogen levels; interaction between allele frequency of the G/A-455 polymorphism, age and smoking. Clin Genet 1996; 50: 184-90.
    PubMedGoogle Scholar
  • 10 Yu S, Sher B, Kudryk B. et al. Intracellular assembly of human fibrinogen. J Biol Chem 1983; 258: 13407-10.
    PubMedGoogle Scholar
  • 11 Roy SN, Mukhopadhyay G, Redman CM. Regulation of fibrinogen assembly. Trans-fection of Hep G2 cells with B beta cDNA specifically enhances synthesis of the three component chains of fibrinogen. J Biol Chem 1990; 265: 6389-93.
    PubMedGoogle Scholar
  • 12 Green FR. Fibrinogen polymorphisms and atherothrombotic disease. Ann N Y Acad Sci 2001; 936: 549-59.
    PubMedGoogle Scholar
  • 13 van’t Hooft FM, von Bahr SJ, Silveira A. et al. Two common, functional polymorphisms in the promoter region of the beta- fibrinogen gene contribute to regulation of plasma fibrinogen concentration. Arterioscler Thromb Vasc Biol 1999; 19: 3063-70.
    CrossrefPubMedGoogle Scholar
  • 14 van der Bom JG. Does plasma fibrinogen increase thrombotic tendency?. Eur J Clin Invest 2002; 32: 721-2.
    CrossrefPubMedGoogle Scholar
  • 15 Tybjaerg-Hansen A, Agerholm-Larsen B, Humphries SE. et al. A common mutation (G-455—>A) in the beta-fibrinogen promoter is an independent predictor of plasma fibrino-gen, but not of ischemic heart disease. A study of 9,127 individuals based on the Copenhagen City Heart Study. J Clin Invest 1997; 99: 3034-9.
    CrossrefPubMedGoogle Scholar
  • 16 Baumann RE, Henschen AH. Linkage disequilibrium relationships among four polymorphisms within the human fibrinogen gene cluster. Hum Genet 1994; 94: 165-70.
    PubMedGoogle Scholar
  • 17 Richards AM, Nicholls MG, Yandle TG. et al. Plasma N-terminal pro-brain natriuretic peptide and adrenomedullin: new neurohormonal predictors of left ventricular function and prognosis after myocardial infarction. Circulation 1998; 97: 1921-9.
    CrossrefPubMedGoogle Scholar
  • 18 Ciulla TA, Sklar RM, Hauser SL. A simple method for DNA purification from peripheral blood. Anal Biochem 1988; 174: 485-8.
    CrossrefPubMedGoogle Scholar
  • 19 Burt MJ, George PM, Upton JD. et al. The significance of haemochromatosis gene mutations in the general population: implications for screening. Gut 1998; 43: 830-6.
    CrossrefPubMedGoogle Scholar
  • 20 Talmud P, Tybjaerg-Hansen A, Bhatnagar D. et al. Rapid screening for specific mutations in patients with a clinical diagnosis of familial hypercholesterolaemia. Atherosclerosis 1991; 89: 137-41.
    CrossrefPubMedGoogle Scholar
  • 21 Baumann RE, Henschen AH. Human fibrino-gen polymorphic site analysis by restriction endonuclease digestion and allele-specific polymerase chain reaction amplification: iden- tification of polymorphisms at positions A alpha 312 and B beta 448. Blood 1993; 82: 2117-24.
    PubMedGoogle Scholar
  • 22 Clauss A. Gerinnungsphysiologische Schnellmethode zur Bestimmung des Fibrinogens. Acta Haematol 1957; 17: 237-46.
    CrossrefPubMedGoogle Scholar
  • 23 Brennan SO, Hammonds B, George PM. Aberrant hepatic processing causes removal of activation peptide and primary polymerisation site from fibrinogen Canterbury (Aα20 Val→Asp). J Clin Invest 1995; 96: 2854-8.
    CrossrefPubMedGoogle Scholar
  • 24 Carr Jr. ME, Hermans J. Size and density of fibrin fibers from turbidity. Macromolecules 1978; 11: 46-50.
    CrossrefPubMedGoogle Scholar
  • 25 Carter AM, Mansfield MW, Stickland MH. et al. Beta-fibrinogen gene-455 G/A polymorphism and fibrinogen levels. Risk factors for coronary artery disease in subjects with NIDDM. Diabetes Care 1996; 19: 1265-8.
    CrossrefPubMedGoogle Scholar
  • 26 Kannel WB, Wolf PA, Castelli WP. et al. Fibrinogen and risk of cardiovascular disease. The Framingham Study. JAMA 1987; 258: 1183-6.
    CrossrefPubMedGoogle Scholar
  • 27 Palmieri V, Celentano A, Roman MJ. et al. Relation of fibrinogen to cardiovascular events is independent of preclinical cardiovascular disease: the Strong Heart Study. Am Heart J 2003; 145: 467-74.
    CrossrefPubMedGoogle Scholar
  • 28 Krobot K, Hense HW, Cremer P. et al. Determinants of plasma fibrinogen: relation to body weight, waist-to- hip ratio, smoking, alcohol, age, and sex. Results from the second MONICA Augsburg survey 1989-1990. Arterioscler Thromb 1992; 12: 780-8.
    CrossrefPubMedGoogle Scholar
  • 29 Hamsten A, Iselius L, De Faire U. et al. Genetic and cultural inheritance of plasma fibrinogen concentration. Lancet 1987; 2: 988-91.
    PubMedGoogle Scholar
  • 30 Humphries SE, Cook M, Dubowitz M. et al. Role of genetic variation at the fibrinogen locus in determination of plasma fibrinogen concentrations. Lancet 1987; 1: 1452-5.
    PubMedGoogle Scholar
  • 31 Brown ET, Fuller GM. Detection of a complex that associates with the Bbeta fibrinogen G-455- A polymorphism. Blood 1998; 92: 3286-93.
    PubMedGoogle Scholar
  • 32 Boekholdt SM, Bijsterveld NR, Moons AH. et al. Genetic variation in coagulation and fibrinolytic proteins and their relation with acute myocardial infarction: a systematic review. Circulation 2001; 104: 3063-8.
    CrossrefPubMedGoogle Scholar
  • 33 Gardemann A, Schwartz O, Haberbosch W. et al. Positive association of the beta fibrinogen H1/H2 gene variation to basal fibrinogen levels and to the increase in fibrinogen concentration during acute phase reaction but not to coronary artery disease and myocardial infarction. Thromb Haemost 1997; 77: 1120-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 34 Renner W, Cichocki L, Forjanics A. et al. G-455A polymorphism of the fibrinogen beta gene and deep vein thrombosis. Eur J Clin Invest 2002; 32: 755-8.
    CrossrefPubMedGoogle Scholar
  • 35 Everse SJ, Spraggon G, Veerapandian L. et al. Crystal structure of fragment double-D from human fibrin with two different bound ligands. Biochemistry 1998; 37: 8637-42.
    CrossrefPubMedGoogle Scholar
  • 36 Everse SJ, Spraggon G, Veerapandian L. et al. Conformational changes in fragments D and double-D from human fibrin(ogen) upon binding the peptide ligand Gly-His-Arg-Pro-amide. Biochemistry 1999; 38: 2941-6.
    CrossrefPubMedGoogle Scholar
  • 37 Villegas A, Hojas R, González FA. et al. Hb Clinico-Madrid II [α90(FG2)Lys>Arg (α1)] and Hb El Escorial [α96(G3)Val>Asp (α2)]: two new α chain variants found during a neonatal period study. Hemoglobin 2003; 27: 185-90.
    CrossrefPubMedGoogle Scholar
  • 38 Lim BC, Ariens RA, Carter AM. et al. Genetic regulation of fibrin structure and function: complex gene- environment interactions may modulate vascular risk. Lancet 2003; 361: 1424-31.
    CrossrefPubMedGoogle Scholar
  • 39 Henschen-Edman AH. Fibrinogen non-inherited heterogeneity and its relationship to function in health and disease. Ann N Y Acad Sci 2001; 936: 580-93.
    PubMedGoogle Scholar
  • 40 Gralnick HR, Givelber H, Abrams E. Dysfibrinogenemia associated with hepatoma. Increased carbohydrate content of the fibrino-gen molecule. N Engl J Med 1978; 299: 221-6.
    CrossrefPubMedGoogle Scholar
  • 41 Dang CV, Shin CK, Bell WR. et al. Fibrinogen sialic acid residues are low affinity calcium-binding sites that influence fibrin assembly. J Biol Chem 1989; 264: 15104-8.
    PubMedGoogle Scholar