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Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM

Righetto, Ricardo D. and Biyani, Nikhil and Kowal, Julia and Chami, Mohamed and Stahlberg, Henning. (2019) Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM. Nature Communications, 10 (1). p. 1722.

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Official URL: https://edoc.unibas.ch/70378/

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Abstract

Electron crystallography can reveal the structure of membrane proteins within 2D crystals under close-to-native conditions. High-resolution structural information can only be reached if crystals are perfectly flat and highly ordered. In practice, such crystals are difficult to obtain. Available image unbending algorithms correct for disorder, but only perform well on images of non-tilted, flat crystals, while out-of-plane distortions are not addressed. Here, we present an approach that employs single-particle refinement procedures to locally unbend crystals in 3D. With this method, density maps of the MloK1 potassium channel with a resolution of 4 Å were obtained from images of 2D crystals that do not diffract beyond 10 Å. Furthermore, 3D classification allowed multiple structures to be resolved, revealing a series of MloK1 conformations within a single 2D crystal. This conformational heterogeneity explains the poor diffraction observed and is related to channel function. The approach is implemented in the FOCUS package.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Stahlberg)
05 Faculty of Science > Departement Biozentrum > Services Biozentrum > BioEM Lab (Chami)
UniBasel Contributors:Stahlberg, Henning and Chami, Mohamed
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Nature Publishing Group
e-ISSN:2041-1723
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:19 Sep 2020 03:10
Deposited On:14 Aug 2020 14:38

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