Abstract |
The serpin plasminogen activator inhibitor-1 (PAI-1) is a potential target for anti-thrombotic 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 IC(50)-values for inactivation of PAI-1 by 4 monoclonal antibodies differed strongly between glycosylated PAI-1 and non-glycosylated 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.
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Authors | Ann Gils, Katrine E Pedersen, Peter Skottrup, Anni Christensen, Dominik Naessens, Johanna Deinum, Jan J Enghild, Paul J Declerck, Peter A Andreasen |
Journal | Thrombosis and haemostasis
(Thromb Haemost)
Vol. 90
Issue 2
Pg. 206-17
(Aug 2003)
ISSN: 0340-6245 [Print] Germany |
PMID | 12888867
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Antibodies, Monoclonal
- Detergents
- Plasminogen Activator Inhibitor 1
- Vitronectin
- Octoxynol
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Topics |
- Antibodies, Monoclonal
(immunology)
- Cell Line
- Detergents
(pharmacology)
- Genetic Variation
- Glycosylation
- Half-Life
- Humans
- Molecular Structure
- Octoxynol
(pharmacology)
- Plasminogen Activator Inhibitor 1
(chemistry, immunology, metabolism)
- Vitronectin
(pharmacology)
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