ジャーナル: J Struct Biol / 年: 2020 タイトル: A microtubule RELION-based pipeline for cryo-EM image processing. 著者: Alexander D Cook / Szymon W Manka / Su Wang / Carolyn A Moores / Joseph Atherton / 要旨: 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.
根拠: microscopy, The assembly is a fragment (repeating unit) of a 13-protofilament GDP-microtubule decorated with NDC-NDC chimera via either of the two N-terminal DC repeats of human doublecortin ...根拠: microscopy, The assembly is a fragment (repeating unit) of a 13-protofilament GDP-microtubule decorated with NDC-NDC chimera via either of the two N-terminal DC repeats of human doublecortin (DCX), as determined by Cryo-EM 3D reconstruction
5-component complex of N-terminal DC domain (NDC) of NDC-NDC chimera (human doublecortin sequence) bound to 13-protofilament GDP-microtubule lattice site composed of 4 tubulin subunits (two alpha-subunits and two beta-subunits)