Capillary electrophoresis determination of the binding affinity of bioactive sulfated polysaccharides to proteins: study of the binding properties of fucoidan to antithrombin

Type Article
Date 2003-04
Language English
Author(s) Varenne A, Gareil P, Colliec-Jouault SylviaORCID, Daniel R
Affiliation(s) Ecole Natl Super Chim Paris, CNRS, UMR 7575, Lab Electrochim & Chim Analyt, F-75231 Paris 05, France.
IFREMER, DRV, VP, BM,Lab Biotechnol & Mol Marines, F-44311 Nantes, France.
Univ Paris 13, CNRS, UMR 7540, Lab Rech Macromol, F-93430 Villetaneuse, France.
Source Analytical Biochemistry (0003-2697) (Elsevier), 2003-04 , Vol. 315 , N. 2 , P. 152-159
DOI 10.1016/S0003-2697(02)00687-5
WOS© Times Cited 44
Keyword(s) Antithrombin, Fucoidan, Electrophoresis, Protein interaction, Polysaccharides
Abstract The interaction of proteins with polysaccharides represents a major and challenging topic in glycobiology, since such complexes mediate fundamental biological mechanisms. An affinity capillary electrophoresis method has been developed to evidence the complex formation and to determine the binding properties between an anticoagulant polysaccharide of marine origin, fucoidan, and a potential target protein, antithrombin. This method is a variant of zonal electrophoresis in the mobility shift format. A fixed amount of protein was injected into a capillary filled with a background electrolyte containing the polysaccharide in varying concentrations. The effective mobility data of the protein were processed according to classical linearization treatments to obtain the binding constant for the polysaccharide/antithrombin complex. The results indicate that fucoidan binds to antithrombin in a 1:1 stoichiometry and with an affinity depending on the molecular weight of the polysaccharide. For heparin, the binding constant obtained similarly is in accordance with the literature. This is the first report showing the implementation of a capillary electrophoresis method contributing to the mechanistic understanding of the biological activities of fucoidan and providing evidence for the complex formation between fucoidan and the protein inhibitor of the coagulation antithrombin. (C) 2003 Elsevier Science (USA). All rights reserved.
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Varenne A, Gareil P, Colliec-Jouault Sylvia, Daniel R (2003). Capillary electrophoresis determination of the binding affinity of bioactive sulfated polysaccharides to proteins: study of the binding properties of fucoidan to antithrombin. Analytical Biochemistry, 315(2), 152-159. Publisher's official version : https://doi.org/10.1016/S0003-2697(02)00687-5 , Open Access version : https://archimer.ifremer.fr/doc/00000/2114/