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![[Figure 2]](zi5010fig2mag.jpg)
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Figure 2
MiCSPARC reconstruction of a KIF5A motor domain-decorated, GMPCPP-stabilized microtubule. (a) Initial rough signal-subtracted reconstruction of a single KIF5A-decorated protofilament (left), which was then used to computationally generate synthetic 3D references for supervised 3D classification (right; colored reconstructions; protofilament number and helical `start' numbers are labeled). (b) The resulting distribution from supervised 3D classification and 3D class `smoothing' of the KIF5A-decorated microtubule dataset using the references in (a). (c) Two views of a final MiCSPARC refinement of a single KIF5A-decorated protofilament after symmetry expansion and α/β-tubulin register correction. Obvious differences in α-tubulin versus β-tubulin S9–S10 loops are indicated. The map was post-processed using EMReady (He et al., 2023  ). (d) Mg-GMPCPP in β-tubulin (stick representation) in the corresponding density (transparent surface representation). (e) Mg-GTP in α-tubulin (stick representation) in the corresponding density (transparent surface representation). (f) AMPPCP in KIF5A (stick representation) in the corresponding density (transparent surface representation). Densities in (d)–(f) are thresholded at the same levels. (g) Seam view of the seam-corrected, 14-protofilament and 3-start microtubule reconstruction obtained using MiCSPARC. The location of the seam is indicated by an arrow. |
![[Figure 2]](zi5010fig2.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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