Identification and characterization of novel lysine-independent apolipoprotein(a)-binding sites in fibrin(ogen) alphaC-domains.

Abstract

Accumulation of lipoprotein(a) (Lp(a)) in atherosclerotic plaques is mediated through interaction of fibrin-(ogen) deposits with the apolipoprotein(a) (apo(a)) moiety of Lp(a). It was suggested that because apo(a) competes with plasminogen for binding to fibrin, causing inhibition of fibrinolysis, it could also promote atherothrombosis. Because the fibrin(ogen) alphaC-domains bind plasminogen and tissue-type plasminogen activator with high affinity in a Lys-dependent manner, we hypothesized that they could also bind apo(a). To test this hypothesis, we studied the interaction between the recombinant apo(a) A10 isoform and the recombinant alphaC-fragment (Aalpha-(221-610)) corresponding to the alphaC-domain by enzyme-linked immunosorbent assay and surface plasmon resonance. Both methods revealed a high affinity interaction (Kd = 19-21 nm) between the immobilized alphaC-fragment and apo(a), indicating that the former contains an apo(a)-binding site. This affinity was comparable to that of apo(a) for fibrin. At the same time, no interaction was observed between soluble fibrinogen and immobilized apo(a), suggesting that, in the former, this and other apo(a)-binding sites are cryptic. Further experiments with truncated recombinant variants of the alphaC-fragment allowed localization of the apo(a)-binding site to the Aalpha-(392-610) region. The presence of epsilon-aminocaproic acid only slightly inhibited binding of apo(a) to the alphaC-fragment, indicating the Lys-independent nature of their interaction. In agreement, the influence of plasminogen or tissue-type plasminogen activator on binding of apo(a) to the alphaC-fragment was minimal. These results indicate that the alphaC-domains contain novel high affinity apo(a)-binding sites that may provide a Lys-independent mechanism for bringing Lp(a) to places of fibrin deposition such as injured vessels or atherosclerotic lesions.

• Authors: Galina Tsurupa, Benoît Ho-Tin-Noé, Eduardo Anglés-Cano, Leonid Medved
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 278 Issue 39 Pg. 37154-9 (Sep 26 2003) ISSN: 0021-9258 [Print] United States
• PMID: 12853452 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Apolipoproteins
• Lipoprotein(a)
• Peptide Fragments
• Fibrin
• Fibrinogen
• Plasminogen
• Apoprotein(a)
• Tissue Plasminogen Activator
• Lysine

Topics

• Apolipoproteins (metabolism)
• Apoprotein(a)
• Binding Sites
• Binding, Competitive
• Fibrin (chemistry, metabolism)
• Fibrinogen (chemistry, metabolism)
• Humans
• Lipoprotein(a) (metabolism)
• Lysine
• Peptide Fragments (metabolism)
• Plasminogen (metabolism)
• Tissue Plasminogen Activator (metabolism)