Abstract
Protein–carbohydrate interactions are often involved in the first step of infection and Pseudomonas aeruginosa produces several proteins that are able to bind specifically to glycan epitopes present on host epithelia. The experimental approaches for studying protein–carbohydrate interaction have been inspired, with some adaptations, from those commonly used for protein–protein or protein–ligand interactions. A range of methods are described herein for detecting lectin activity, screening for monosaccharide or oligosaccharide specificity, determining the affinity of binding together with thermodynamics and kinetics parameters, and producing crystal of lectin–carbohydrate complexes for further structural studies.
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Acknowledgments
The authors acknowledge funding from ANR grants PA-Antiadh (ANR-09-JCJC-0047) and Glycoasterix (ANR-08-PCVI-0028). Support from GDR Pseudomonas and Association Vaincre la Mucoviscidose is also acknowledged.
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de Bentzmann, S., Varrot, A., Imberty, A. (2014). Monitoring Lectin Interactions with Carbohydrates. In: Filloux, A., Ramos, JL. (eds) Pseudomonas Methods and Protocols. Methods in Molecular Biology, vol 1149. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-0473-0_32
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DOI: https://doi.org/10.1007/978-1-4939-0473-0_32
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