Quantifying CBM-Carbohydrate Interactions Using Microscale Thermophoresis

Haiyang Wu; Cédric Y Montanier; Claire Dumon

doi:10.1007/978-1-0716-3151-5_7

Quantifying CBM-Carbohydrate Interactions Using Microscale Thermophoresis

Methods Mol Biol. 2023:2657:103-114. doi: 10.1007/978-1-0716-3151-5_7.

Authors

Haiyang Wu¹, Cédric Y Montanier¹, Claire Dumon²

Affiliations

¹ TBI, Université de Toulouse, CNRS, INRAE, INSA, Toulouse, France.
² TBI, Université de Toulouse, CNRS, INRAE, INSA, Toulouse, France. claire.dumon@insa-toulouse.fr.

PMID: 37149525
DOI: 10.1007/978-1-0716-3151-5_7

Abstract

Microscale thermophoresis (MST) is an emerging technology for studying a broad range of biomolecular interactions with a high sensitivity. The affinity constant can be obtained for a wide range of molecules within minutes based on reactions in microliters. Here we describe the application of MST in quantifying protein-carbohydrate interactions. A CBM3a and a CBM4 are titrated with insoluble substrate (cellulose nanocrystal) and soluble oligosaccharide (xylohexaose), respectively.

Keywords: Binding studies; Fluorescence quenching; Fluorescent label; Kd (dissociation constant); Microscale thermophoresis (MST).

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Carbohydrates*
Cellulose*
Protein Binding

Substances

Carbohydrates
Cellulose