title: Antifibrinolytic effect of recombinant apolipoprotein(a) in vitro is primarily due to attenuation of tPA-mediated Glu-plasminogen activation.
abstract: The effect of a 17-kringle form of recombinant apo(a) [r-apo(a)] on in vitro fibrin clot lysis was studied. In these assays, fibrin clots were formed in the wells of microtiter plates, and lysis of the clots was monitored by measurement of the turbidity at 405 nm. The results indicate that r-apo(a) produces a dose-dependent antifibrinolytic effect in clots formed using either purified components or barium-adsorbed plasma. This effect was found to be independent of clot structure, since lysis of clots formed using both high and low concentrations of thrombin was prolonged by r-apo(a) to the same extent. The two components of the antifibrinolytic effect of r-apo(a) were determined to be (i) attenuation of tPA-mediated plasminogen activation (the major component) and (ii) inhibition of plasmin degradation of fibrin, although r-apo(a) did not directly attenuate plasmin activity, as measured by S-2251 hydrolysis. r-Apo(a) interfered most substantially with tPA-mediated activation of Glu-plasminogen and less substantially with tPA-mediated Lys-plasminogen activation and urokinase-mediated activation of plasminogen. In summary, we have demonstrated that apo(a) is able to attenuate fibrin clot lysis in vitro, primarily as a consequence of the interference by apo(a) with tPA-mediated Glu-plasminogen activation. These studies illuminate possible mechanisms by which Lp(a) may contribute to the development of vascular disease in vivo.
authors.affiliation: Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
authors.forename: W
authors.initials: W
authors.lastname: Sangrar
authors.forename: L
authors.initials: L
authors.lastname: Bajzar
authors.forename: M E
authors.initials: ME
authors.lastname: Nesheim
authors.forename: M L
authors.initials: ML
authors.lastname: Koschinsky
mesh_terms: D000595:Amino Acid Sequence; D000933:Antifibrinolytic Agents / Q000494:pharmacology*; D001054:Apolipoproteins A / Q000494:pharmacology*; D005341:Fibrinolysin / Q000037:antagonists & inhibitors; D006801:Humans; D066298:In Vitro Techniques; D018082:Kringles; D017270:Lipoprotein(a) / Q000502:physiology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000037:antagonists & inhibitors / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism*; D011994:Recombinant Proteins / Q000494:pharmacology; D010959:Tissue Plasminogen Activator / Q000037:antagonists & inhibitors*
chemical_list: D000933:Antifibrinolytic Agents; D001054:Apolipoproteins A; D017270:Lipoprotein(a); D010446:Peptide Fragments; D011994:Recombinant Proteins; C033809:lys-plasmin; C018998:lysyl-plasminogen; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin