title: Myosin: a noncovalent stabilizer of fibrin in the process of clot dissolution.
abstract: Myosin modulates the fibrinolytic process as a cofactor of the tissue plasminogen activator and as a substrate of plasmin. We report now that myosin is present in arterial thrombi and it forms reversible noncovalent complexes with fibrinogen and fibrin with equilibrium dissociation constants in the micromolar range (1.70 and 0.94 microM, respectively). Competition studies using a peptide inhibitor of fibrin polymerization (glycl-prolyl-arginyl-proline [GPRP]) indicate that myosin interacts with domains common in fibrinogen and fibrin and this interaction is independent of the GPRP-binding polymerization site in the fibrinogen molecule. An association rate constant of 1.81 x 10(2) M(-1) x s(-1) and a dissociation rate constant of 3.07 x 10(-4) s(-1) are determined for the fibrinogen-myosin interaction. Surface plasmon resonance studies indicate that fibrin serves as a matrix core for myosin aggregation. The fibrin clots equilibrated with myosin are stabilized against dissolution initiated by plasminogen and tissue-type plasminogen activator (tPA) or urokinase (at fibrin monomer-myosin molar ratio as high as 30) and by plasmin under static and flow conditions (at fibrin monomer-myosin molar ratio lower than 15). Myosin exerts similar effects on the tPA-induced dissolution of blood plasma clots. Covalent modification involving factor XIIIa does not contribute to this stabilizing effect; myosin is not covalently attached to the clot by the time of complete cross-linking of fibrin. Thus, our in vitro data suggest that myosin detected in arterial thrombi binds to the polymerized fibrin, in the bound form its tPA-cofactor properties are masked, and the myosin fibrin clot is relatively resistant to plasmin.
authors.affiliation: Department of Medical Biochemistry, Semmelweis University, Budapest, Hungary.
authors.forename: Krasimir
authors.initials: K
authors.lastname: Kolev
authors.forename: Kiril
authors.initials: K
authors.lastname: Tenekedjiev
authors.forename: Katalin
authors.initials: K
authors.lastname: Ajtai
authors.forename: Ilona
authors.initials: I
authors.lastname: Kovalszky
authors.forename: Judit
authors.initials: J
authors.lastname: Gombas
authors.forename: Balázs
authors.initials: B
authors.lastname: Váradi
authors.forename: Raymund
authors.initials: R
authors.lastname: Machovich
mesh_terms: D001665:Binding Sites; D001798:Blood Proteins / Q000494:pharmacology; D019281:Dimerization; D005337:Fibrin / Q000378:metabolism*; D005342:Fibrinolysis; D006801:Humans; D007700:Kinetics; D009218:Myosins / Q000378:metabolism / Q000502:physiology*; D010447:Peptide Hydrolases / Q000378:metabolism; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013927:Thrombosis; D010959:Tissue Plasminogen Activator / Q000494:pharmacology
chemical_list: D001798:Blood Proteins; D005337:Fibrin; D010447:Peptide Hydrolases; D010959:Tissue Plasminogen Activator; D009218:Myosins