Selective Interface Detection:Mapping Binding Site Contacts in Membrane Proteins by NMR Spectroscopy

摘要

Interactions with proteins are at the root of all living systems.In large molecules,such interactions usually involve only a small fraction of the total molecular surface area,for example,the ligand binding pocket of a large receptor protein.NMR methods for selective observation of intermolecular contact surfaces can simplify spectra while targeting a region of biological interest.Here,we propose a method for selective observation of interfacial 3-7 A contacts between isotopically ~(13)C-labeled and unlabeled moieties by solid state magic angle spinning(MAS)NMR.We demonstrate this new technique in an application to an unlabeled membrane-embedded G-protein coupled receptor(GPCR),rhodopsin(Rho),containing the uniformly ~(13)C-labeled ligand,11-cis-retinal.