Routine sub-2.5 Å cryo-EM structure determination of GPCRs

Radostin Danev (), Matthew Belousoff, Yi-Lynn Liang, Xin Zhang, Fabian Eisenstein, Denise Wootten and Patrick M. Sexton
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Radostin Danev: University of Tokyo
Matthew Belousoff: Monash University
Yi-Lynn Liang: Monash University
Xin Zhang: Monash University
Fabian Eisenstein: University of Tokyo
Denise Wootten: Monash University
Patrick M. Sexton: Monash University

Nature Communications, 2021, vol. 12, issue 1, 1-10

Abstract: Abstract Cryo-electron microscopy (cryo-EM) of small membrane proteins, such as G protein-coupled receptors (GPCRs), remains challenging. Pushing the performance boundaries of the technique requires quantitative knowledge about the contribution of multiple factors. Here, we present an in-depth analysis and optimization of the main experimental parameters in cryo-EM. We combined actual structural studies with methods development to quantify the effects of the Volta phase plate, zero-loss energy filtering, objective lens aperture, defocus magnitude, total exposure, and grid type. By using this information to carefully maximize the experimental performance, it is now possible to routinely determine GPCR structures at resolutions better than 2.5 Å. The improved fidelity of such maps enables the building of better atomic models and will be crucial for the future expansion of cryo-EM into the structure-based drug design domain. The optimization guidelines given here are not limited to GPCRs and can be applied directly to other small proteins.

Date: 2021
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Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-24650-3

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DOI: 10.1038/s41467-021-24650-3

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