通过核磁共振光谱法表征蛋白质 - 碳水化合物相互作用

Characterization of Protein-Carbohydrate Interactions by NMR Spectroscopy.

作者信息

Grondin Julie M, Langelaan David N, Smith Steven P

机构信息

Lethbridge Research Center, Agriculture and Agri-Food Canada, 5403-1 Ave. South, 3000, Lethbridge, AB, Canada, T1J 4P4.

Department of Biochemistry and Molecular Biology, Dalhousie University, Halifax, NS, Canada, B3H 4R2.

出版信息

Methods Mol Biol. 2017;1588:143-156. doi: 10.1007/978-1-4939-6899-2_11.

DOI:10.1007/978-1-4939-6899-2_11

PMID:28417366

Abstract

Solution-state nuclear magnetic resonance (NMR) spectroscopy can be used to monitor protein-carbohydrate interactions. Two-dimensional H-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 ) of any identified interactions, and to map the carbohydrate binding site on the structure of the 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.

摘要

溶液状态核磁共振（NMR）光谱可用于监测蛋白质与碳水化合物的相互作用。本章所述的基于二维H-N异核单量子相干（HSQC）的技术可快速有效地筛选出一组可能的碳水化合物结合伴侣，量化任何已鉴定相互作用的解离常数（K），并在蛋白质结构上绘制碳水化合物结合位点。在此，我们描述了用单糖N-乙酰半乳糖胺（GalNAc）滴定产气荚膜梭菌（CpCBM32）的32家族碳水化合物结合模块，其中我们计算了相互作用的表观解离，并将GalNAc结合位点映射到CpCBM32的结构上。