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Title | A microtubule RELION-based pipeline for cryo-EM image processing. |
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Journal, issue, pages | J Struct Biol, Vol. 209, Issue 1, Page 107402, Year 2020 |
Publish date | Jan 1, 2020 |
![]() | Alexander D Cook / Szymon W Manka / Su Wang / Carolyn A Moores / Joseph Atherton / ![]() |
PubMed 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 ...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. |
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Methods | EM (single particle) / EM (helical sym.) |
Resolution | 3.8 - 4.5 Å |
Structure data | ![]() EMDB-10131: ![]() EMDB-10195: |
Chemicals | ![]() ChemComp-GDP: ![]() ChemComp-GTP: ![]() ChemComp-MG: |
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