Type of Publication: | Journal article |
Publication status: | Published |
URI (citable link): | http://nbn-resolving.de/urn:nbn:de:bsz:352-2-14ggsx2jn8tfh6 |
Author: | Köhn, Birgit; Ponnandai Shanmugavel, Kumaravel; Wu, Min; Kovermann, Michael; Wittung-Stafshede, Pernilla |
Year of publication: | 2018 |
Published in: | Biophysical journal ; 115 (2018), 6. - pp. 1007-1018. - ISSN 0006-3495. - eISSN 1542-0086 |
Pubmed ID: | 30173886 |
DOI (citable link): | https://dx.doi.org/10.1016/j.bpj.2018.07.040 |
Summary: |
The copper-transporting ATPase ATP7B is essential for loading of copper ions to copper-dependent enzymes in the secretory pathway; its inactivation results in Wilson disease. In contrast to copper-ion uptake by the cytoplasmic domains, ATP7B-mediated copper-ion release in the Golgi has not been explored yet. We demonstrate here that a luminal loop in ATP7B, rich in histidine/methionine residues, binds reduced copper (Cu(I)) ions, and identified copper-binding residues play an essential role in ATP7B-mediated metal ion release. NMR experiments on short-peptide models demonstrate that three methionine and two histidine residues are specifically involved in Cu(I) ion binding; with these residues replaced by alanines, no Cu(I) ion interaction is detected. Although more than one Cu(I) ion can interact with the wild-type peptide, removing either all histidine or all methionine residues reduces the stoichiometry to one Cu(I) ion binding per peptide. Using a yeast complementation assay, we show that for efficient copper transport by full-length ATP7B, the complete set of histidine and methionine residues in the lumen loop are required. The replacement of histidine/methionine residues by alanines does not perturb overall ATP7B structure, as the localization of ATP7B variants in yeast cells matches that of the wild-type protein. Thus, in similarity to ATP7A, ATP7B also appears to have a luminal "exit" copper ion site.
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Subject (DDC): | 540 Chemistry |
Link to License: | In Copyright |
Bibliography of Konstanz: | Yes |
Refereed: | Yes |
KÖHN, Birgit, Kumaravel PONNANDAI SHANMUGAVEL, Min WU, Michael KOVERMANN, Pernilla WITTUNG-STAFSHEDE, 2018. A Luminal Loop of Wilson Disease Protein Binds Copper and Is Required for Protein Activity. In: Biophysical journal. 115(6), pp. 1007-1018. ISSN 0006-3495. eISSN 1542-0086. Available under: doi: 10.1016/j.bpj.2018.07.040
@article{Kohn2018-09-18Lumin-44419, title={A Luminal Loop of Wilson Disease Protein Binds Copper and Is Required for Protein Activity}, year={2018}, doi={10.1016/j.bpj.2018.07.040}, number={6}, volume={115}, issn={0006-3495}, journal={Biophysical journal}, pages={1007--1018}, author={Köhn, Birgit and Ponnandai Shanmugavel, Kumaravel and Wu, Min and Kovermann, Michael and Wittung-Stafshede, Pernilla} }
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