Qualitative and Quantitative Characterization of Protein-Carbohydrate Interactions by NMR Spectroscopy.

Autor: Grondin JM; Department of Education, Simon Fraser University, Burnaby, AB, Canada., Langelaan DN; Department of Biochemistry & Molecular Biology, Dalhousie University, Halifax, NS, Canada., Smith SP; Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON, Canada. steven.smith@queensu.ca.
Jazyk: angličtina
Zdroj: Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2657, pp. 115-128.
DOI: 10.1007/978-1-0716-3151-5_8
Abstrakt: Solution-state nuclear magnetic resonance (NMR) spectroscopy can be used to monitor protein-carbohydrate interactions. Two-dimensional 1 H- 15 N heteronuclear single quantum coherence (HSQC)-based techniques described in this chapter can be used quickly and effectively to screen a set of possible carbohydrate-binding partners, to quantify the dissociation constant (K d ) of any identified interactions, and to the map the carbohydrate-binding site on the structure of a protein. Here, we describe the titration of a family 32 carbohydrate-binding module from Clostridium perfringens (CpCBM32) with the monosaccharide N-acetylgalactosamine (GalNAc), in which we calculate the apparent dissociation of the interaction and map the GalNAc binding site onto the structure of CpCBM32. This approach can be applied to other CBM- and protein-ligand systems.
(© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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