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 using affinity electrophoresis (AEP) is described. Polyacrylamide gels covalently conjugated with 2-acetamido-2-deoxy-D-glucopyranose (GlcNAc) residues were successfully prepared by radical copolymerization of highly reactive 3-(N-acryloylamino)propyl 2-acetamido-2-deoxy-beta-D-glucopyranoside with acrylamide in the presence of N,N'-methylenebisacrylamide (BIS). When the glycogels carrying various densities of GlcNAc branches were employed for polyacrylamide gel electrophoresis (PAGE) of lectins, the mobilities of wheat germ agglutinin (WGA) were specifically 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 of WGA with immobilized GlcNAc residue can be determined by combined use of this stable glycogel and an automated gel-scanning system associated with fluorometric spectroscopy. The association constant of WGA with the GlcNAc moiety was estimated to be 1.24 x 10(4) M-1.
Ohmae
M
M
Division of Bio-Science, Hokkaido University, Sapporo.
Suzuki
S
S
Tokura
S
S
Nishi
N
N
Nishimura
S I
SI
D050260:Carbohydrate Metabolism; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005782:Gels / Q000737:chemistry; D037102:Lectins / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy
D005782:Gels; D037102:Lectins