SYNTHETIC GLYCOGELS FOR AFFINITY ELECTROPHORESIS - A FACILE AND EFFICIENT METHOD FOR INVESTIGATING SUGAR-LECTIN INTERACTION

A novel and efficient method for analyzing sugar-lectin interaction us ing affinity electrophoresis (AEP) is described, Polyacrylamide gels c ovalently conjugated with 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) residues were successfully prepared by radical copolymerization of hi ghly reactive 3-(N-acryloylamino)propyl 2-acetamido-2-deoxy-beta-D-glu copyranoside with acrylamide in the presence of N,N-methylenebisacryla mide (BIS), When the glycogels carrying various densities of GlcNAc br anches were employed for polyacrylamide gel electrophoresis (PAGE) of lectins, the mobilities of wheat germ agglutinin (WGA) were specifical ly reduced by increasing the concentrations of the GlcNAc residues in gels, although concanavalin A (Con A) showed no significant change in the mobility, It was also demonstrated that the association constant o f WGA with immobilized GlcNAc residue can be determined by combined us e of this stable glycogel and an automated gel-scanning system associa ted with fluorometric spectroscopy, The association constant of WGA wi th the GlcNAc moiety was estimated to be 1.24X10(4) M(-1).