title: NMR analysis of carbohydrate-binding interactions in solution: an approach using analysis of saturation transfer difference NMR spectroscopy.
abstract: One of the most commonly used ligand-based NMR methods for detecting ligand binding is saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy. The STD NMR method is an invaluable technique for assessing carbohydrate-lectin interactions in solution, because STD NMR can be used to detect weak ligand binding (Kd ca. 10(-3)-10(-8) M). STD NMR spectra identify the binding epitope of a carbohydrate ligand when bound to lectin. Further, the STD NMR method uses 1H-detected NMR spectra of only the carbohydrate, and so only small quantities of non-labeled lectin are required. In this chapter, I describe a protocol for the STD NMR method, including the experimental procedures used to acquire, process, and analyze STD NMR data, using STD NMR studies for methyl-β-D-galactopyranoside (β-Me-Gal) binding to the C-terminal domain of an R-type lectin from earthworm (EW29Ch) as an example.
authors.affiliation: National Food Research Institute, National Agriculture and Food Research Organization (NARO), 2-1-12, Kannondai, Tsukuba, 305-8642, Ibaraki, Japan, hemmi@affrc.go.jp.
authors.forename: Hikaru
authors.initials: H
authors.lastname: Hemmi
mesh_terms: D053000:Analytic Sample Preparation Methods; D000818:Animals; D037102:Lectins / Q000737:chemistry* / Q000378:metabolism*; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008756:Methylgalactosides / Q000737:chemistry* / Q000378:metabolism*; D009835:Oligochaeta; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012996:Solutions
chemical_list: D037102:Lectins; D008024:Ligands; D008756:Methylgalactosides; D012996:Solutions; C031552:methyl beta-galactoside