Affinity Electrophoresis for Analysis of Catalytic Module-Carbohydrate Interactions

Abstract

Affinity electrophoresis has long been used to study the interaction between proteins and large soluble ligands. The technique has been found to have great utility for the examination of polysaccharide binding by proteins, particularly carbohydrate-binding modules (CBMs). In recent years carbohydrate surface binding sites of proteins, mostly enzymes, have also been investigated by this method. Here we describe a protocol for identifying binding interactions between enzyme catalytic modules and a variety of carbohydrate ligands.

Keywords: Affinity electrophoresis; Carbohydrate-binding module; Dissociation constant; Polyacrylamide gel electrophoresis; Polysaccharide; Surface binding site.