title: Saturation transfer difference NMR spectroscopy as a technique to investigate protein-carbohydrate interactions in solution.
abstract: Saturation transfer difference (STD) Nuclear Magnetic Resonance (NMR) spectroscopy is a powerful method for studying protein-ligand interactions in solution. The STD NMR method is capable of identifying the binding epitope of a ligand when bound to its receptor protein. Ligand protons that are in close contact with the receptor protein receive a higher degree of saturation, and as a result stronger STD NMR signals can be observed. Protons that are either less or not involved in the binding process reveal no STD NMR signals. Therefore, the STD NMR method is an excellent tool to investigate how a binding ligand interacts with its receptor molecule. The STD NMR experiment is easy to implement and only small amounts of native protein are required. This chapter comprises a detailed experimental protocol to acquire STD NMR spectra and determine the binding epitope of a ligand bound to its target protein. As representative examples the ligands uridyl-triphosphate (UTP) and uridyl-glucose-diphosphate (UDP-glucose) when bound to the Leishmania major UDP-glucose-pyrophosphorylase (UGP) as target protein are examined.
authors.affiliation: Institute for Glycomics, Griffith University, Gold Coast Campus, Gold Coast, QLD, 4222, Australia. t.haselhorst@griffith.edu.au
authors.forename: Thomas
authors.initials: T
authors.lastname: Haselhorst
authors.forename: Anne-Christin
authors.initials: AC
authors.lastname: Lamerz
authors.forename: Mark von
authors.initials: Mv
authors.lastname: Itzstein
mesh_terms: D000818:Animals; D001665:Binding Sites; D050260:Carbohydrate Metabolism / Q000502:physiology*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis / Q000737:chemistry; D006801:Humans; D008954:Models, Biological; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012996:Solutions / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D002241:Carbohydrates; D011506:Proteins; D012996:Solutions