Solid-State NMR Approaches to Study Protein Structure and Protein-Lipid Interactions.
Fiche publication
Date publication
janvier 2019
Journal
Methods in molecular biology (Clifton, N.J.)
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BECHINGER Burkhard
Tous les auteurs :
Aisenbrey C, Salnikov ES, Raya J, Michalek M, Bechinger B
Lien Pubmed
Résumé
Solid-state NMR spectroscopy has been developed for the investigation of membrane-associated polypeptides and remains one of the few techniques to reveal high-resolution structural information in liquid-disordered phospholipid bilayers. In particular, oriented samples have been used to investigate the structure, dynamics and topology of membrane polypeptides. Much of the previous solid-state NMR work has been developed and performed on peptides but the technique is constantly expanding towards larger membrane proteins. Here, a number of protocols are presented describing among other the reconstitution of membrane proteins into oriented membranes, monitoring membrane alignment by P solid-state NMR spectroscopy, investigations of the protein by one- and two-dimensional N solid-state NMR and measurements of the lipid order parameters using H solid-state NMR spectroscopy. Using such methods solid-state NMR spectroscopy has revealed a detailed picture of the ensemble of both lipids and proteins and their mutual interdependence in the bilayer environment.
Mots clés
Bicelle, Channel, Cross-polarization, Detergent, Helix topology, Lee–Goldburg decoupling, Membrane protein structure, Membrane reconstitution, Oriented bilayer, PISEMA, Protein insertion, Separated local field spectroscopy, Surface alignment, Transmembrane orientation
Référence
Methods Mol. Biol.. 2019 ;2003:563-598