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Archives of cardiovascular diseases
Volume 102, n° S1
page 6 (mars 2009)
Doi : 10.1016/S1875-2136(09)72134-5
Themes et posters

A001 Affinity of low molecular weight fucoidan for P-selectin triggers its binding to activated human platelets
 

L. Bachelet-Violette 1, I. Bertholon 1, D. Lavigne 1, R. Vassy 2, M. Jandrot-Perrus 1, F. Chaubet 1, D. Letourneur 1
1 Inserm U698, Paris, France 
2 CNRS UMR 7033, Bobigny, France 

A — Atherosclerose, hemostase, inflammation, age

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Background

P-selectin is an adhesion receptor expressed on activated platelets and endothelial cells. Its natural ligand, P-selectin glycoprotein ligand-1, is expressed on leucocytes and the P-selectin/PSGL-1 interaction is involved in leukocyte rolling. We have compared the interaction of P-selectin with several low molecular weight polysaccharides: fucoidan, heparin and dextran sulfate.

Methods

Binding assays were obtained from the interaction of the polysaccharides with Sialyl Lewis X and PSGL-1 based constructs onto microtiter plates coated with P-selectin. SELDI TOF mass spectrometry was performed with anionic chips arrays coated with P-selectin in the absence or in the presence of polysaccharides. Kd were obtained from surface plasmon resonance experiments with immobilized Pselectin constructs, polysaccharides being injected in the mobile phase. Human whole blood flow cytometry experiments were performed with fluorescein isothiocyanate labelled polysaccharides with or without platelets activators.

Results

The fucoidan prevented P-selectin binding to Sialyl Lewis X with an IC50 of 20nM as compared to 400nM for heparin and >25000nM for dextran sulfate. It exhibited the highest affinity for immobilized Pselectin with a KD of 1.2nM, two orders of magnitude greater than the KD of the other polysaccharides. Mass spectrometry evidenced the formation of a complex between P-selectin and fucoidan. The intensity of the fucoidan binding to platelets was dependent on the level of platelet activation. Competition between fucoidan and an anti P-selectin antibody demonstrated the specificity of the interaction.

General significance

Low molecular weight fucoidan is a promising therapeutic agent of natural origin for biomedical applications.

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Affinity of low molecular weight fucoidan for P-selectin triggers its binding to activated human platelets.

Bachelet L, Bertholon I, Lavigne D, Vassy R, Jandrot-Perrus M, Chaubet F, Letourneur D.

Biochim Biophys Acta. 2008 Nov 5.


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