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. Association constants determined by AE were in excellent agreement with values obtained by isothermal titration calorimetry and fluorescence titration. The method was adapted to the identification, study and characterization of mutant 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.
Tomme
P
Protein Engineering Network of Centres of Excellence, Department of Microbiology and Immunology and The Biotechnology Laboratory, #237-6174 University of British Columbia, V6T 1Z3, Vancouver, BC, Canada
Boraston
A
Kormos
JM
Warren
RA
Kilburn
DG