Multiple binding with identical linkage: a mechanism that explains the effect of lipoprotein(a) on fibrinolysis.
We have previously shown that both recombinant apo(a) and native Lp(a) inhibit the binding of Glu-plasminogen to fibrin surfaces [Fleury & Anglés-Cano (1991) Biochemistry 30, 7630-7638; Rouy et al. (1992) Biochemistry 31, 6332-6339]. The aim of the present study was to characterize the mechanism of this inhibition and to define the parameters governing binding when two different Lp(a) species compete with plasminogen for fibrin, a situation that may be found in vivo in subjects heterozygous for the apo(a) trait. The Kd for the binding of plasminogen to fibrin was 660 nM whereas the affinity of Lp(a) was inversely related to apo(a) size (Kd range: 50 to > 500 nM). To determine the effect of plasminogen on Lp(a) binding and reciprocally, competition experiments were performed. The Kd of either Lp(a) or plasminogen for fibrin remained unchanged in the presence of the other competitor whereas Bmax, the maximal amount bound, was importantly decreased. In a similar fashion, competition for fibrin binding among Lp(a) isoforms was shown with the use of Lp(a) density fractions containing varying proportions of isoforms B (approximately 460 kDa) and S3 (approximately 640 kDa); variations in Kd values (from 141 nM to 460 nM) as a function of the relative content in isoform S3 were observed. Altogether, these results are indicative of multiple binding by ligands that bind with different affinities to equivalent but independent sites. Thus, in plasma from heterozygous subjects, the influence of each Lp(a) isoform on fibrinolysis will depend on their affinity for fibrin and on their concentration relative to each other and to plasminogen.
Hervio
L
L
Institut National de la Santé et de la Recherche Médicale, INSERM U.143, Centre Hospitalier Universitaire de Bicêtre, Paris, France.
Durlach
V
V
Girard-Globa
A
A
Anglés-Cano
E
E
D001667:Binding, Competitive; D005337:Fibrin / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D005342:Fibrinolysis / Q000187:drug effects*; D006801:Humans; D007700:Kinetics; D017270:Lipoprotein(a) / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology*; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology
D017270:Lipoprotein(a); D011994:Recombinant Proteins; D005337:Fibrin; D005340:Fibrinogen; D010958:Plasminogen