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TitleRole of surfactants in electron cryo-microscopy film preparation.
Journal, issue, pagesBiophys J, Vol. 122, Issue 10, Page 1846-1857, Year 2023
Publish dateMay 16, 2023
AuthorsBaptiste Michon / Uriel López-Sánchez / Jéril Degrouard / Hugues Nury / Amélie Leforestier / Emmanuelle Rio / Anniina Salonen / Manuela Zoonens /
PubMed AbstractSingle-particle electron cryo-microscopy (cryo-EM) has become an effective and straightforward approach to determine the structure of membrane proteins. However, obtaining cryo-EM grids of sufficient ...Single-particle electron cryo-microscopy (cryo-EM) has become an effective and straightforward approach to determine the structure of membrane proteins. 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-HT receptor whose structure has been solved in APol. These findings may speed up the process of grid optimization to obtain high-resolution structures of membrane proteins.
External linksBiophys J / PubMed:37077048 / PubMed Central
MethodsEM (single particle)
Resolution3.4 Å
Structure data

EMDB-15471: Mouse serotonin 5-HT3A receptor in amphipols A8-35
Method: EM (single particle) / Resolution: 3.4 Å

Source
  • Mus musculus (house mouse)

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