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. 2021 Oct 26;12:6173. doi: 10.1038/s41467-021-26427-0

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a The model depicts the work-flow of sample preparation for cryo-EM: MonoUb-Cyclin B1-NT was incubated briefly with purified 20S proteasomes at 1:5 (proteasome:substrate) molar ratio on ice, frozen within 5 min, and then vitrified for further cryo-EM single-particle analysis. b Difference map rendering the symmetric pattern between the trans- and cis-rings of the 20S-alone map (upper) and 20S + monoUb-Cyclin B1-NT map (lower). The difference map (red density) was generated by using a 180° rotated (around the pseudo twofold axis) map minus its original unrotated map (transparent gray). c The top view of density maps from the cis α-ring of 20S only (blue) and the S1 class of 20S + monoUb-CyclinB11-NT (brick red) dataset. d Comparison of the 20S-alone (transparent blue) and the S1 class of 20S + MonoUb-Cyclin B1-NT (brick red) maps in the two β-rings focused on the β-annulus regions. Note the density loss in some of the annulus loops or β-sheet regions exposed to or close to the catalytic chamber in both β-rings.