Journal
ENZYME AND MICROBIAL TECHNOLOGY
Volume 27, Issue 7, Pages 453-458Publisher
ELSEVIER SCIENCE INC
DOI: 10.1016/S0141-0229(00)00246-5
Keywords
affinity electrophoresis; competition; carbohydrate-binding module; CBM; cellulose-binding domain; polysaccharide
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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. (C) 2000 Elsevier Science Inc. All rights reserved.
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