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

Pengxin Chai, Qinhui Rao, Yue Wang, Kai Zhang

Methods in molecular biology (Clifton, N.J.) 2023

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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. © 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Pengxin Chai, Qinhui Rao, Yue Wang, Kai Zhang. High-Resolution Structural Analysis of Dyneins by Cryo-electron Microscopy. Methods in molecular biology (Clifton, N.J.). 2023;2623:257-279