Affinity electrophoresis for the identification and characterization of soluble sugar binding by carbohydrate-binding modules

Affinity electrophoresis was used to identify and quantify the interaction of carbohydrate-binding modules (CBMs) with soluble polysaccharides. Associ ation constants determined by AE were in excellent agreement with values ob tained by isothermal titration calorimetry and fluorescence titration. The method was adapted to the identification, study and characterization of mut ant carbohydrate-binding modules with altered affinities and specificities. Competition affinity electrophoresis was used to monitor binding of small, soluble mono- and disaccharides to one of the modules. (C) 2000 Elsevier S cience Inc. All rights reserved.