Role of surfactants in electron cryo-microscopy film preparation - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Biophysical Journal Année : 2023

Role of surfactants in electron cryo-microscopy film preparation

Résumé

Single-particle electron cryo-microscopy (cryo-EM) has become an effective and straightforward approach to determine the structure of membrane proteins (MPs). However, obtaining cryo-EM grids of sufficient quality for high-resolution structural analysis remains a major bottleneck. One of the difficulties arises from the presence of detergents, which often leads to a lack of control of the ice thickness. Amphipathic polymers such as amphipols (APols) are detergent substitutes, which have proven to be valuable tools for cryo-EM studies. In this work, we investigate the physico-chemical behavior of APol- and detergent-containing solutions and show a correlation with the properties of vitreous thin films in cryo-EM grids. This study provides new insight on the potential of APols, allowing a better control of ice thickness while limiting protein adsorption at the air-water interface, as shown with the full-length mouse serotonin 5-HT3A receptor whose structure has been solved in APol. These findings may speed up the process of grid optimization to obtain high-resolution structures of MPs.
Fichier principal
Vignette du fichier
Michon_preprint.pdf (13.89 Mo) Télécharger le fichier
Origine : Fichiers produits par l'(les) auteur(s)

Dates et versions

hal-04271843 , version 1 (06-11-2023)

Identifiants

Citer

Baptiste Michon, Uriel López-Sánchez, Jéril Degrouard, Hugues Nury, Amélie Leforestier, et al.. Role of surfactants in electron cryo-microscopy film preparation. Biophysical Journal, 2023, 122 (10), pp.1846-1857. ⟨10.1016/j.bpj.2023.04.016⟩. ⟨hal-04271843⟩
27 Consultations
17 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More