High-performance affinity chromatography for the purification of heparin-binding proteins from detergent-solubilized smooth muscle cell membranes

Journal of Chromatography. B, Biomedical Sciences and Applications
A S ClairboisJ Jozefonvicz

Abstract

Heparin and heparan sulfates are regulators of cellular events including adhesion, proliferation and migration. In particular, the antiproliferative effect of heparin on smooth muscle cell (SMC) growth is well described. However, its mechanism of action remains unclear. Numerous results suggest an endocytosis mediated by a still unknown heparin receptor on vascular SMCs. In order to identify a putative heparin receptor on SMCs that could be involved in heparin signalling, affinity chromatography supports were developed. In this paper, we describe high-performance liquid affinity chromatography (HPLAC) supports obtained from silica beads coated with dextran polymer substituted by a calculated amount of diethylaminoethyl functions. With a polysaccharide dextran layer, this type of support can be grafted with specific ligands, such as heparin, using conventional coupling methods. In a previous work, we demonstrated, using butanedioldiglycidyl ether, that silica stationary phases coupled to heparin could be used for the fast elution and good peak resolution of heparin-binding proteins. In the present work, an affinity chromatographic fraction of SMC membrane extracts was analyzed by sodium dodecyl sulfate-polyacrylamide gel electro...Continue Reading

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Citations

Mar 4, 2000·Journal of Controlled Release : Official Journal of the Controlled Release Society·D LetourneurM Cansell
Dec 24, 2011·Journal of the American Chemical Society·Mingfei ShaoXue Duan
Jul 6, 2000·Infection Control and Hospital Epidemiology : the Official Journal of the Society of Hospital Epidemiologists of America·B Montdargent, D Letourneur
Mar 15, 2001·Journal of Pharmaceutical and Biomedical Analysis·M M Rhemrev-BoomM Raedts

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