High-Resolution Structural Analysis of Dyneins by Cryo-electron Microscopy.

Methods Mol Biol

Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, CT, USA.

Published: January 2023

Cryo-electron microscopy (cryo-EM) has become the mainstream technique for studying macromolecular structures. Determining the structures of protein complexes is more accessible to structural biologists than ever before. Nevertheless, obtaining high-resolution structures of molecular motors like dynein is still an extremely challenging goal due to their troublesome behaviors in ice, their exceedingly flexible conformations, and their intricate architectures. Dynein is a large molecular machine that drives the movement of many essential cellular cargos and is also the key force generator that powers ciliary motility. High-resolution structural information of dyneins in different states is critical for the in-depth mechanistic understanding of their roles in cells. Here, we summarize the cryo-EM approaches that we have used to study the structures of outer-arm dynein arrays bound to microtubule doublets. Our approaches can be applied to other similar structures and further optimized to deal with even more complicated targets.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10371436PMC
http://dx.doi.org/10.1007/978-1-0716-2958-1_16DOI Listing

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