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J. Biol. Chem., Vol. 277, Issue 52, 50245-50254, December 27, 2002

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Carbohydrate-binding Modules Recognize Fine Substructures of Cellulose^*

Bradley W. McLean^§¶, Alisdair B. Boraston^§¶**, Darren Brouwer, Nooshafarin Sanaie^§§, Colin A. Fyfe, R. Antony J. Warren^§, Douglas G. Kilburn^§¶, and Charles A. Haynes^¶§§¶¶

From the  Protein Engineering Network of Centres of Excellence, PENCE Inc., National Business Centre, Edmonton, Alberta T6G 2S2, Canada, ^§ Department of Microbiology and Immunology, ^¶ The Biotechnology Laboratory, the  Department of Chemistry, and the ^§§ Department of Chemical and Biological Engineering, University of British Columbia, Vancouver, British Columbia V6T 1Z4, Canada

Competition isotherms are used to identify the set of cellulose substructures to which cellulose binding modules (CBMs) from families 2a, 3, 4, 9, and 17 bind. The experiments are based on coupling a unique fluorescent tag to each CBM in a manner that does not alter the natural binding properties of the CBM and therefore allows the surface and solution concentrations of each CBM to be monitored as a function of time and composition. Adsorption and surface exchange of like or competing CBMs are monitored using a range of cellulose preparations varying in both crystallinity and provenance. CBMs from families 2a, 3, 4, 9, and 17 are shown to recognize different physical forms of prepared cellulose. The demonstration of the very fine binding specificity of cellulose-specific CBMs implies that the polysaccharide targets of CBMs extend down to the resolution of cellulose microstructures.

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