Study of lectin-ganglioside interactions by high-performance liquid affinity chromatography.
A high-performance affinity column containing immobilized modified GM1 (lyso-GM1) was used to study the binding of an endogenous human brain lectin (HBL) in comparison with other carbohydrate-binding proteins. The proteins are previously converted into biotinylated derivatives. Detection of biotinylated proteins in the eluates by a microtitre plate assay ensures good sensitivity. The maximum binding capacity of the adsorbent for HBL is obtained in Tris buffer supplemented with beta-mercaptoethanol. The binding is inhibitable by specific sugar. It is concluded that the use of immobilized glycolipids in analytical high-performance liquid affinity chromatographic methods may serve as models in the study of interactions between gangliosides and carbohydrate-binding proteins.
Caron
M
M
Laboratoire de Biochimie et Technologie des Protéines, Université Paris-Nord, France.
Joubert-Caron
R
R
Cartier
J R
JR
Chadli
A
A
Bladier
D
D
D001710:Biotin / Q000737:chemistry; D001923:Brain Chemistry; D002772:Cholera Toxin / Q000737:chemistry; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005677:G(M1) Ganglioside / Q000737:chemistry; D005732:Gangliosides / Q000737:chemistry*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry
D005732:Gangliosides; D037102:Lectins; D037121:Plant Lectins; D005677:G(M1) Ganglioside; D001710:Biotin; D002772:Cholera Toxin