Cook, A.D. and Manka, Szymon W. and Wang, S. and Moores, Carolyn A. and Atherton, Joe (2020) A microtubule RELION-based pipeline for cryo-EM image processing. Journal of Structural Biology 209 (1), p. 107402. ISSN 1047-8477.
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Abstract
Microtubules are polar filaments built from αβ-tubulin heterodimers that exhibit a range of architectures in vitro and in vivo. Tubulin heterodimers are arranged helically in the microtubule wall but many physiologically relevant architectures exhibit a break in helical symmetry known as the seam. Noisy 2D cryo-electron microscopy projection images of pseudo-helical microtubules therefore depict distinct but highly similar views owing to the high structural similarity of α- and β-tubulin. The determination of the αβ-tubulin register and seam location during image processing is essential for alignment accuracy that enables determination of biologically relevant structures. Here we present a pipeline designed for image processing and high-resolution reconstruction of cryo-electron microscopy microtubule datasets, based in the popular and user-friendly RELION image-processing package, Microtubule RELION-based Pipeline (MiRP). The pipeline uses a combination of supervised classification and prior knowledge about geometric lattice constraints in microtubules to accurately determine microtubule architecture and seam location. The presented method is fast and semi-automated, producing near-atomic resolution reconstructions with test datasets that contain a range of microtubule architectures and binding proteins.
Metadata
| Item Type: | Article |
|---|---|
| Keyword(s) / Subject(s): | Microtubule, pseudo-helical symmetry, cryo-EM, RELION, 3D reconstruction |
| School: | Birkbeck Faculties and Schools > Faculty of Science > School of Natural Sciences |
| Depositing User: | Administrator |
| Date Deposited: | 16 Oct 2019 11:19 |
| Last Modified: | 14 Mar 2025 19:05 |
| URI: | https://eprints.bbk.ac.uk/id/eprint/29525 |
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