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. 2024 Mar 27;15:2690. doi: 10.1038/s41467-024-47001-4

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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/.

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Fig. 4 — Supporting cryo-EM density is shown in Supplementary Figs. 8–12 and 22. A In the E1 state (HMA4^apo) the first of the CPC cysteines (C597 of M4) is oriented away, preventing copper uptake. This configuration is stabilized by Y912, N913 (M5) and S945 (M6). B In the E1-Cu state (HMA4^Cu) an entry site for copper is formed, in-between M352 of M1 as well as C597 and C599. C Comparison of the copper uptake region in the E2P state (HMA4^AlF). Note that both the two CPC cysteines are directed away from M352 and hence are incompatible with copper uptake. D The equivalent E1-Cu state as in panel b for AfCopA. E Copper release in the E2P state (HMA4^AlF). An opening from the surrounding lumen reaches the CPC cysteines, and it is lined by numerous sulfur-containing residues of MA, M2, and M6, including the conserved M295, M391, and M941 thereby permitting copper release (Supplementary Fig. 1a). The MA link to the soluble domains may represent a point of regulating ion release. Blue and red surfaces denote positively and negatively charged cavities.