title: Factor XIII A-Subunit V34L Variant Affects Thrombus Cross-Linking in a Murine Model of Thrombosis.
abstract: OBJECTIVE
  Factor XIII (FXIII) cross-links fibrin upon activation by thrombin. Activation involves cleavage at residue 37 by thrombin, releasing an activation peptide. A common polymorphism (valine to leucine variant at residue 34, V34L), located in the activation peptide, has been associated with increased activation rates and paradoxically a protective effect in cardiovascular disease. There is, currently, no data available on the effects of V34L from in vivo models of thrombosis. We examined the effect of FXIII V34L on clot formation and cross-linking in vivo.
  
  
  RESULTS
  We generated a panel of full-length recombinant human FXIII-A2 variants with amino acid substitutions in the activation peptide to investigate the effect of these variants on activation rate, and we used wild-type, V34L, and alanine to glycine variant at residue 33 variants to study the effects of varying FXIII activation rate on thrombus formation in a murine model of FeCl3 injury. FXIII activation assay showed that residues 29, 30, 33, and 34 play a critical role in thrombin interaction. Full-length recombinant human FXIII-A2 V34L has significant effects on clot formation, structure, and lysis in vitro, using turbidity assay. This variant influenced fibrin cross-linking but not size of the thrombus in vivo.
  
  
  CONCLUSIONS
  Mutations in the activation peptide of full-length recombinant FXIII regulate activation rates by thrombin, and V34L influences in vivo thrombus formation by increased cross-linking of the clot.
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.).
authors.forename: Cédric
authors.initials: C
authors.lastname: Duval
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.).
authors.forename: Majid
authors.initials: M
authors.lastname: Ali
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.).
authors.forename: Waleed W
authors.initials: WW
authors.lastname: Chaudhry
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.).
authors.forename: Victoria C
authors.initials: VC
authors.lastname: Ridger
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.). R.A.S.Ariens@leeds.ac.uk.
authors.forename: Robert A S
authors.initials: RA
authors.lastname: Ariëns
authors.affiliation: From the Thrombosis and Tissue Repair Group, Division of Cardiovascular and Diabetes Research, Multidisciplinary Cardiovascular Research Centre, Leeds Institute of Cardiovascular And Metabolic Medicine, Faculty of Medicine and Health, University of Leeds, Leeds, United Kingdom (C.D., M.A., W.W.C., R.A.S.A., H.P.); and Department of Cardiovascular Science, Faculty of Medicine, Dentistry, and Health, University of Sheffield, Sheffield, South Yorkshire, United Kingdom (V.C.R.).
authors.forename: Helen
authors.initials: H
authors.lastname: Philippou
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D001777:Blood Coagulation; D001780:Blood Coagulation Tests; D004195:Disease Models, Animal; D005177:Factor XIII Deficiency / Q000097:blood / Q000235:genetics / Q000378:metabolism; D026122:Factor XIIIa / Q000008:administration & dosage / Q000235:genetics / Q000378:metabolism*; D005337:Fibrin / Q000378:metabolism*; D005838:Genotype; D006801:Humans; D008297:Male; D057507:Mice, 129 Strain; D008808:Mice, Inbred CBA; D018345:Mice, Knockout; D009154:Mutation; D010641:Phenotype; D011994:Recombinant Proteins / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism*; D013997:Time Factors; D020246:Venous Thrombosis / Q000097:blood / Q000235:genetics / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D005337:Fibrin; D026122:Factor XIIIa; D013917:Thrombin
keywords: factor XIII; fibrinogen; mutation; thrombin; venous thrombosis