Biochemical importance of glycosylation of plasminogen activator inhibitor-1

Ann Gils; Katrine E. Pedersen; Peter Skottrup; Anni Christensen; Dominik Naessens; Johanna Deinum; Jan J. Enghild; Paul J. Declerck; Peter A. Andreasen

doi:10.1160/TH03-01-0034

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 2003; 90(02): 206-217
DOI: 10.1160/TH03-01-0034

Blood Coagulation, Fibrinolysis and Cellular Haemostasis

Schattauer GmbH

Biochemical importance of glycosylation of plasminogen activator inhibitor-1

Ann Gils^*

¹Laboratory for Pharmaceutical Biology and Phytopharmacology, Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, Belgium

,

Katrine E. Pedersen^*

²Department of Molecular Biology, Aarhus University, Denmark

,

Peter Skottrup^*

²Department of Molecular Biology, Aarhus University, Denmark

,

Anni Christensen

²Department of Molecular Biology, Aarhus University, Denmark

,

Dominik Naessens

¹Laboratory for Pharmaceutical Biology and Phytopharmacology, Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, Belgium

,

Johanna Deinum

³AstraZeneca, R&D, Mölndal, Sweden

,

Jan J. Enghild

²Department of Molecular Biology, Aarhus University, Denmark

,

Paul J. Declerck

¹Laboratory for Pharmaceutical Biology and Phytopharmacology, Faculty of Pharmaceutical Sciences, Katholieke Universiteit Leuven, Belgium

,

Peter A. Andreasen

²Department of Molecular Biology, Aarhus University, Denmark

› Institutsangaben

Abstract

Summary

The serpin plasminogen activator inhibitor-1 (PAI-1) is a potential target for anti-thombotic and anti-cancer therapy. PAI-1 has 3 potential sites for N-linked glycosylation. We demonstrate here that PAI-1 expressed recombinantly or naturally by human cell lines display a heterogeneous glycosylation pattern of the sites at N209 and N265, while that at N329 is not utilised. The IC₅₀-values for inactivation of PAI-1 by 4 monoclonal antibodies differed strongly between glycosylated PAI-1 and non-glycosy-lated PAI-1 expressed in E. coli. For 3 antibodies, an overlap of the epitopes with the glycosylation sites could be excluded as explanation for the differential reactivity. The latency transition of non-glycosylated, but not of glycosylated PAI-1, was strongly accelerated by a non-ionic detergent. The different biochemical properties of glycosylated and non-glycosylated PAI-1 depended specifically on glycosylation of either one or the other of the utilised sites. The PAI-1-binding protein vitronectin reversed the changes associated with the lack of glycosylation at one of the sites. Our results stress the importance of the source of PAI-1 when studying the mechanisms of action of PAI-1-inactivating compounds of potential clinical importance.

Keywords

Thombosis - arteriosclerosis - metastasis - serpin - vitronectin

Volltext

Referenzen

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