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. 2024 Jan 2;43(2):250–276. doi: 10.1038/s44318-023-00018-9

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© The Author(s) 2024

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. Creative Commons Public Domain Dedication waiver http://creativecommons.org/publicdomain/zero/1.0/ applies to the data associated with this article, unless otherwise stated in a credit line to the data, but does not extend to the graphical or creative elements of illustrations, charts, or figures. This waiver removes legal barriers to the re-use and mining of research data. According to standard scholarly practice, it is recommended to provide appropriate citation and attribution whenever technically possible.

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Figure EV2 — (A) AlphaFold models of UBA6 and UBA7 and the crystal structures of UBA1, UBA2 and UBA3 are shown. The structures were aligned and electrostatic potential was calculated as described in the methods. The yellow arrows indicate the location of the groove within the SCCH domains. UBA6, UBA1 and UBA7 form an extended lobe within the SCCH, which is missing in UBA2 and UBA3. The groove in UBA7 is covered and do not exist in UBA2 and UBA3. The grooves in UBA1 and UBA6 are highly similar in terms of structure but present significantly different electro potential surfaces. The gradient from negative (red) to positive (blue) charge is shown. The figure was prepared by PyMol. (B) Microscale thermophoresis interaction analysis of cy5-7Ala-peptide against UBA6 or UBA1 SCCH domain. The dose-response curve of cy5-7Ala-peptide titrated against increasing concentrations of the SCCH domain is presented. Results are mean ± s.e.m. n = 4 biological replicates, Two-way ANOVA, Sidak’s test. ****P < 0.0001.