Retrieving high-resolution information from disordered 2D crystals by single-particle cryo-EM. View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2019-12

AUTHORS

Ricardo D Righetto, Nikhil Biyani, Julia Kowal, Mohamed Chami, Henning Stahlberg

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. More... »

PAGES

1722

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/s41467-019-09661-5

DOI

http://dx.doi.org/10.1038/s41467-019-09661-5

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1113380250

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30979902


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