journal menu
![[Figure 2]](jb5071fig2mag.jpg)
|
|
Figure 2
Cryo-EM identification and high-resolution structure determination of ArnA contaminant. (a) Representative motion-corrected cryo-EM micrograph. Cryo-EM screening revealed heterogeneous fields containing regions of aggregation alongside dispersed, homogeneous particles suitable for single-particle analysis. White arrows mark dispersed ArnA particles. (b) 2D classification of cryo-EM particles. Scale bar: 11 nm. Unexpectedly, 2D classification yielded highly homogeneous class averages exhibiting threefold-symmetric features characteristic of the ArnA hexamer, rather than views consistent with the intended KIF17–IFT70 complex. (c) Cryo-EM reconstruction of ArnA obtained from the dataset. Ab initio reconstruction and subsequent refinement produced a threefold-symmetric map corresponding to the ArnA hexamer, refined to an overall resolution of 3.23 Å. The resulting density is consistent with previously reported ArnA architectures, confirming that the dominant particles in the dataset correspond to ArnA rather than the intended target complex. The final reconstructed ArnA map and local resolution distribution are shown. (d) Comparison of the ArnA map obtained in this study with that reported by Caliseki and coworkers. (e) Superposition of the ArnA structural model determined here with the corresponding EM map, shown in transparent gray. (f) Structural superimposition of the ArnA models from this study and from that of Caliseki and coworkers. |
![[Figure 2]](jb5071fig2.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
access
