Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH
Roles of fibrin α- and γ-chain specific cross-linking by FXIIIa in fibrin structure and function
Cédric Duval
1
Theme Thrombosis, Division of Cardiovascular and Diabetes Research, Leeds Institute of Genetics, Health and Therapeutics and Multidisciplinary Cardiovascular Research Centre, Faculty of Medicine and Health, University of Leeds, UK
,
Peter Allan
1
Theme Thrombosis, Division of Cardiovascular and Diabetes Research, Leeds Institute of Genetics, Health and Therapeutics and Multidisciplinary Cardiovascular Research Centre, Faculty of Medicine and Health, University of Leeds, UK
2
Molecular and Nanoscale Physics Group, School of Physics and Astronomy, University of Leeds, UK
,
Simon D. A. Connell
2
Molecular and Nanoscale Physics Group, School of Physics and Astronomy, University of Leeds, UK
,
Victoria C. Ridger
3
Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, UK
,
Helen Philippou
1
Theme Thrombosis, Division of Cardiovascular and Diabetes Research, Leeds Institute of Genetics, Health and Therapeutics and Multidisciplinary Cardiovascular Research Centre, Faculty of Medicine and Health, University of Leeds, UK
,
Robert A. S. Ariëns
1
Theme Thrombosis, Division of Cardiovascular and Diabetes Research, Leeds Institute of Genetics, Health and Therapeutics and Multidisciplinary Cardiovascular Research Centre, Faculty of Medicine and Health, University of Leeds, UK
› Author AffiliationsFinancial support: This study was supported by Medical Research Council (G0901546) and British Heart Foundation (RG/13/2/30104) grants.
Factor XIII is responsible for the cross-linking of fibrin γ-chains in the early stages of clot formation, whilst α-chain cross-linking occurs at a slower rate. Although γ- and α-chain cross-linking was previously shown to contribute to clot stiffness, the role of cross-linking of both chains in determining clot structure is currently unknown. Therefore, the aim of this study was to determine the role of individual α- and γ-chain cross-linking during clot formation, and its effects on clot structure. We made use of a recombinant fibrinogen (γQ398N/Q399N/K406R), which does not allow for y-chain cross-linking. In the absence of cross-linking, intact D-D interface was shown to play a potential role in fibre appearance time, clot stiffness and elasticity. Cross-linking of the fibrin α-chain played a role in the thickening of the fibrin fibres over time, and decreased lysis rate in the absence of α2-antiplasmin. We also showed that α-chain cross-linking played a role in the timing of fibre appearance, straightening fibres, increasing clot stiffness and reducing clot deformation. Cross-linking of the γ-chain played a role in fibrin fibre appearance time and fibre density. Our results show that α- and γ-chain cross-linking play independent and specific roles in fibrin clot formation and structure.
3
Purves L,
Purves M,
Brandt W.
Cleavage of fibrin-derived D-dimer into monomers by endopeptidase from puff adder venom (Bitis arietans) acting at cross-linked sites of the gamma-chain. Sequence of carboxy-terminal cyanogen bromide gamma-chain fragments. Biochemistry 1987; 26: 4640-4646.
4
Cottrell BA,
Strong DD,
Watt KW.
et al. Amino acid sequence studies on the alpha chain of human fibrinogen. Exact location of cross-linking acceptor sites. Biochemistry 1979; 18: 5405-5410.
5
Matsuka YV,
Medved LV,
Migliorini MM.
et al. Factor XIIIa-catalyzed cross-linking of recombinant alpha C fragments of human fibrinogen. Biochemistry 1996; 35: 5810-5816.
8
Shen L,
Lorand L.
Contribution of fibrin stabilisation to clot strength. Supplementation of factor XIII-deficient plasma with the purified zymogen. J Clin Invest 1983; 71: 1336-1341.
9
Ryan EA,
Mockros LF,
Stern AM.
et al. Influence of a natural and a synthetic inhibitor of factor XIIIa on fibrin clot rheology. Biophys J 1999; 77: 2827-2836.
11
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: 988-995.
12
Undas A,
Ariens RA.
Fibrin clot structure and function: a role in the pathophysiology of arterial and venous thromboembolic diseases. Arterioscler Thromb Vasc Biol 2011; 31: e88-e99.
13
Standeven KF,
Carter AM,
Grant PJ.
et al. Functional analysis of fibrin gamma-chain cross-linking by activated factor XIII: determination of a cross-linking pattern that maximizes clot stiffness. Blood 2007; 110: 902-907.
14
Lord ST,
Strickland E,
Jayjock E.
Strategy for recombinant multichain protein synthesis: fibrinogen B beta-chain variants as thrombin substrates. Biochemistry 1996; 35: 2342-2348.
15
Takebe M,
Soe G,
Kohno I.
et al. Calcium ion-dependent monoclonal antibody against human fibrinogen: preparation, characterisation, and application to fibrinogen purification. Thromb Haemost 1995; 73: 662-667.
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.
17
Abou-Saleh RH,
Connell SD,
Harrand R.
et al. Nanoscale probing reveals that reduced stiffness of clots from fibrinogen lacking 42 N-terminal Bbeta-chain residues is due to the formation of abnormal oligomers. Biophys J 2009; 96: 2415-2427.
18
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.
19
Evans RM,
Tassieri M,
Auhl D.
et al. Direct conversion of rheological compliance measurements into storage and loss moduli. Phys Rev E Stat Nonlin Soft Matter Phys 2009; 80: 012501.
20
Shen LL,
McDonagh RP,
McDonagh J.
et al. Fibrin gel structure: influence of calcium and covalent cross-linking on the elasticity. Biochem Biophys Res Commun 1974; 56: 793-798.
21
Spraggon G,
Everse SJ,
Doolittle RF.
Crystal structures of fragment D from human fibrinogen and its crosslinked counterpart from fibrin. Nature 1997; 389: 455-462.
22
Collet JP,
Moen JL,
Veklich YI.
et al. The alphaC domains of fibrinogen affect the structure of the fibrin clot, its physical properties, and its susceptibility to fibrinolysis. Blood 2005; 106: 3824-3830.
23
Chernysh IN,
Nagaswami C,
Purohit PK.
et al. Fibrin clots are equilibrium polymers that can be remodelled without proteolytic digestion. Sci Rep 2012; 02: 879.
26
Francis CW,
Marder VJ.
Increased resistance to plasmic degradation of fibrin with highly crosslinked alpha-polymer chains formed at high factor XIII concentrations. Blood 1988; 7195: 1361-1365.
27
Fraser SR,
Booth NA,
Mutch NJ.
The antifibrinolytic function of factor XIII is exclusively expressed through alpha(2)-antiplasmin cross-linking. Blood 2011; 117: 6371-6374.