title: The mutation in fibrinogen Bicêtre II (gamma Asn308-->Lys) does not affect the binding of t-PA and plasminogen to fibrin.
abstract: The aim of this study was to investigate the interactions of t-PA and plasminogen with fibrin derived from an abnormal fibrinogen detected in a 40-year-old male patient who had had an episode of thrombophlebitis with pulmonary embolism. An abnormal fibrinogen was diagnosed on the basis of prolonged thrombin and reptilase times also detected in two other family members. Fibrinogen purified from plasma, in the presence of protease inhibitors, by glycine precipitations, gel filtration and affinity chromatography, was devoid of plasminogen, fibronectin, and vWf. SDS-PAGE analysis according to Laemmli under reducing conditions, showed an abnormal gamma chain (approximately 50% of the total) migrating in a more anodic position (M(r) 48 kDa). By PCR amplification and DNA sequencing, the abnormality was identified as an Asn308-->Lys mutation of the gamma chain. Since such a mutation constitutes a new plasmin cleavage site as first reported for fibrinogen Kyoto I, it may modify interactions of plasminogen and t-PA with carboxy-terminal lysine residues. Ligand-binding studies were therefore performed using intact and plasmin-degraded fibrin surfaces obtained from the abnormal fibrinogen. The plasminogen and t-PA binding isotherms obtained with the abnormal fibrinogen were similar to the control. Moreover, the stimulation by fibrin of plasminogen activation by t-PA was not different from the control. These results suggest (i) that the lysine 308 residue may not be exposed to plasmin cleavage in fibrin, and (ii) that the thrombotic accident of the propositus cannot be explained by an abnormality of the plasminogen/t-PA binding to fibrin.
authors.affiliation: Unité 143, Institut National de la Santé et de la Recherche Médicale, Centre Hospitalo-Universitaire de Bicêtre, France.
authors.forename: P
authors.initials: P
authors.lastname: Grailhe
authors.forename: C
authors.initials: C
authors.lastname: Boyer-Neumann
authors.forename: F
authors.initials: F
authors.lastname: Haverkate
authors.forename: J
authors.initials: J
authors.lastname: Grimbergen
authors.forename: M J
authors.initials: MJ
authors.lastname: Larrieu
authors.forename: E
authors.initials: E
authors.lastname: Anglés-Cano
mesh_terms: D000328:Adult; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D005337:Fibrin / Q000378:metabolism*; D015241:Fibrinogens, Abnormal / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006487:Hemostasis; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D010375:Pedigree; D010958:Plasminogen / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011655:Pulmonary Embolism / Q000097:blood / Q000235:genetics; D017422:Sequence Analysis, DNA; D013918:Thrombin Time; D013924:Thrombophlebitis / Q000097:blood* / Q000235:genetics; D010959:Tissue Plasminogen Activator / Q000378:metabolism*
chemical_list: D015241:Fibrinogens, Abnormal; C085517:fibrinogen Bicetre II; D005337:Fibrin; D010958:Plasminogen; D010959:Tissue Plasminogen Activator