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Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner
The vacuolar ATPase (V‐ATPase) is a rotary motor proton pump that is regulated by an assembly equilibrium between active holoenzyme and autoinhibited V(1)‐ATPase and V(o) proton channel subcomplexes. Here, we report cryo‐EM structures of yeast V‐ATPase assembled in vitro from lipid nanodisc reconsti...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8804929/ https://www.ncbi.nlm.nih.gov/pubmed/34918374 http://dx.doi.org/10.15252/embj.2021109360 |
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author | Khan, Md. Murad Lee, Seowon Couoh‐Cardel, Sergio Oot, Rebecca A Kim, Hyunmin Wilkens, Stephan Roh, Soung‐Hun |
author_facet | Khan, Md. Murad Lee, Seowon Couoh‐Cardel, Sergio Oot, Rebecca A Kim, Hyunmin Wilkens, Stephan Roh, Soung‐Hun |
author_sort | Khan, Md. Murad |
collection | PubMed |
description | The vacuolar ATPase (V‐ATPase) is a rotary motor proton pump that is regulated by an assembly equilibrium between active holoenzyme and autoinhibited V(1)‐ATPase and V(o) proton channel subcomplexes. Here, we report cryo‐EM structures of yeast V‐ATPase assembled in vitro from lipid nanodisc reconstituted V(o) and mutant V(1). Our analysis identified holoenzymes in three active rotary states, indicating that binding of V(1) to V(o) provides sufficient free energy to overcome V(o) autoinhibition. Moreover, the structures suggest that the unequal spacing of V(o)’s proton‐carrying glutamic acid residues serves to alleviate the symmetry mismatch between V(1) and V(o) motors, a notion that is supported by mutagenesis experiments. We also uncover a structure of free V(1) bound to Oxr1, a conserved but poorly characterized factor involved in the oxidative stress response. Biochemical experiments show that Oxr1 inhibits V(1)‐ATPase and causes disassembly of the holoenzyme, suggesting that Oxr1 plays a direct role in V‐ATPase regulation. |
format | Online Article Text |
id | pubmed-8804929 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-88049292022-02-11 Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner Khan, Md. Murad Lee, Seowon Couoh‐Cardel, Sergio Oot, Rebecca A Kim, Hyunmin Wilkens, Stephan Roh, Soung‐Hun EMBO J Articles The vacuolar ATPase (V‐ATPase) is a rotary motor proton pump that is regulated by an assembly equilibrium between active holoenzyme and autoinhibited V(1)‐ATPase and V(o) proton channel subcomplexes. Here, we report cryo‐EM structures of yeast V‐ATPase assembled in vitro from lipid nanodisc reconstituted V(o) and mutant V(1). Our analysis identified holoenzymes in three active rotary states, indicating that binding of V(1) to V(o) provides sufficient free energy to overcome V(o) autoinhibition. Moreover, the structures suggest that the unequal spacing of V(o)’s proton‐carrying glutamic acid residues serves to alleviate the symmetry mismatch between V(1) and V(o) motors, a notion that is supported by mutagenesis experiments. We also uncover a structure of free V(1) bound to Oxr1, a conserved but poorly characterized factor involved in the oxidative stress response. Biochemical experiments show that Oxr1 inhibits V(1)‐ATPase and causes disassembly of the holoenzyme, suggesting that Oxr1 plays a direct role in V‐ATPase regulation. John Wiley and Sons Inc. 2021-12-17 2022-02-01 /pmc/articles/PMC8804929/ /pubmed/34918374 http://dx.doi.org/10.15252/embj.2021109360 Text en © 2021 The Authors Published under the terms of the CC BY NC ND 4.0 license https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Articles Khan, Md. Murad Lee, Seowon Couoh‐Cardel, Sergio Oot, Rebecca A Kim, Hyunmin Wilkens, Stephan Roh, Soung‐Hun Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title | Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title_full | Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title_fullStr | Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title_full_unstemmed | Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title_short | Oxidative stress protein Oxr1 promotes V‐ATPase holoenzyme disassembly in catalytic activity‐independent manner |
title_sort | oxidative stress protein oxr1 promotes v‐atpase holoenzyme disassembly in catalytic activity‐independent manner |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8804929/ https://www.ncbi.nlm.nih.gov/pubmed/34918374 http://dx.doi.org/10.15252/embj.2021109360 |
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