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Thromb Haemost 2001; 86(05): 1229-1237
DOI: 10.1055/s-0037-1616056
Review Article
Schattauer GmbH

The Kringle V-Protease Domain Is a Fibrinogen Binding Region within Apo(a)

Song Xue
1   The Scripps Research Institute, Department of Vascular Biology, La Jolla, California, USA
,
Edwin L. Madison   *
1   The Scripps Research Institute, Department of Vascular Biology, La Jolla, California, USA
,
Lindsey A. Miles
1   The Scripps Research Institute, Department of Vascular Biology, La Jolla, California, USA
› InstitutsangabenThis work was supported in part by The National Institutes of Health, Grant HL38272, HL45934, HL31950 and by Grant-In-Aid #9650636 from the American Heart Association. Dr. Xue is the recipient of an Individual National Research Service Award HL09721.
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Publikationsverlauf

Received 22. August 2000

Accepted after revision 04. Juni 2001

Publikationsdatum:
13. Dezember 2017 (online)

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Summary

Lp(a) binds directly to fibrin and competes for the interaction of plasminogen with this substrate. This competition may play a role in the proatherothrombogenic consequences of high Lp(a) levels. Previous studies by us and others showed that apo(a) Kringle IV-10 competes for the interaction of Lp(a) with plasmin-treated fibrinogen. However, kringle IV-10 cannot account for the entire high affinity interaction of Lp(a) with fibrinogen. Therefore, we tested the hypothesis that the apo(a) kringle V protease-like domain (KV-PD) could interact with plasmin-treated fibrinogen. We cloned the apo(a) KV-PD region from a human liver cDNA library. Fusion apo(a) KV-PD was expressed in COS 7 cells and purified from the conditioned media. Western blotting of the apo(a) KV-PD protein revealed two bands migrating with apparent molecular weights of 45K and 48K. When fusion apo(a) KV-PD was treated with O-glycosidase and neuraminidase, the higher molecular weight band disappeared suggesting that the apo(a) KV-PD was O-glycosylated. Apo(a) KV-PD bound to plasmin-treated fibrinogen in a dose-dependent fashion. An EC50 of 3.9 ± 0.2 μM was determined for this interaction. Treatment of the apo(a) KV-PD with O-glycosidase did not significantly affect its ability to bind to plasmin-treated fibrinogen. In addition, apo(a) KV-PD competed for the binding of 125I-Lp(a) to plasmin-treated fibrinogen. An IC50 of 7.90 ± 0.95 μM was obtained. Our data suggest that the KV-PD of apo(a) shares binding sites on plasmin-treated fibrinogen with Lp(a) and also may participate in the interaction of the Lp(a) particle with plasmin-treated fibrinogen.

Keywords

Apo(a) - kringle - fibrinogen

* Dr. Madison’s present address is: Edwin L. Madison, Ph. D., Director, Molecular Biology, Corvas International, Inc., 3030 Science Park Road, San Diego, CA 92121


 

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