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Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes
Chloroplast F(o)F(1)-ATP synthase (CF(o)CF(1)) converts proton motive force into chemical energy during photosynthesis. Although many studies have been done to elucidate the catalytic reaction and its regulatory mechanisms, biochemical analyses using the CF(o)CF(1) complex have been limited because...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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National Academy of Sciences
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9964038/ https://www.ncbi.nlm.nih.gov/pubmed/36716358 http://dx.doi.org/10.1073/pnas.2218187120 |
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author | Akiyama, Kentaro Ozawa, Shin-Ichiro Takahashi, Yuichiro Yoshida, Keisuke Suzuki, Toshiharu Kondo, Kumiko Wakabayashi, Ken-ichi Hisabori, Toru |
author_facet | Akiyama, Kentaro Ozawa, Shin-Ichiro Takahashi, Yuichiro Yoshida, Keisuke Suzuki, Toshiharu Kondo, Kumiko Wakabayashi, Ken-ichi Hisabori, Toru |
author_sort | Akiyama, Kentaro |
collection | PubMed |
description | Chloroplast F(o)F(1)-ATP synthase (CF(o)CF(1)) converts proton motive force into chemical energy during photosynthesis. Although many studies have been done to elucidate the catalytic reaction and its regulatory mechanisms, biochemical analyses using the CF(o)CF(1) complex have been limited because of various technical barriers, such as the difficulty in generating mutants and a low purification efficiency from spinach chloroplasts. By taking advantage of the powerful genetics available in the unicellular green alga Chlamydomonas reinhardtii, we analyzed the ATP synthesis reaction and its regulation in CF(o)CF(1). The domains in the γ subunit involved in the redox regulation of CF(o)CF(1) were mutated based on the reported structure. An in vivo analysis of strains harboring these mutations revealed the structural determinants of the redox response during the light/dark transitions. In addition, we established a half day purification method for the entire CF(o)CF(1) complex from C. reinhardtii and subsequently examined ATP synthesis activity by the acid–base transition method. We found that truncation of the β-hairpin domain resulted in a loss of redox regulation of ATP synthesis (i.e., constitutively active state) despite retaining redox-sensitive Cys residues. In contrast, truncation of the redox loop domain containing the Cys residues resulted in a marked decrease in the activity. Based on this mutation analysis, we propose a model of redox regulation of the ATP synthesis reaction by the cooperative function of the β-hairpin and the redox loop domains specific to CF(o)CF(1). |
format | Online Article Text |
id | pubmed-9964038 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | National Academy of Sciences |
record_format | MEDLINE/PubMed |
spelling | pubmed-99640382023-07-30 Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes Akiyama, Kentaro Ozawa, Shin-Ichiro Takahashi, Yuichiro Yoshida, Keisuke Suzuki, Toshiharu Kondo, Kumiko Wakabayashi, Ken-ichi Hisabori, Toru Proc Natl Acad Sci U S A Biological Sciences Chloroplast F(o)F(1)-ATP synthase (CF(o)CF(1)) converts proton motive force into chemical energy during photosynthesis. Although many studies have been done to elucidate the catalytic reaction and its regulatory mechanisms, biochemical analyses using the CF(o)CF(1) complex have been limited because of various technical barriers, such as the difficulty in generating mutants and a low purification efficiency from spinach chloroplasts. By taking advantage of the powerful genetics available in the unicellular green alga Chlamydomonas reinhardtii, we analyzed the ATP synthesis reaction and its regulation in CF(o)CF(1). The domains in the γ subunit involved in the redox regulation of CF(o)CF(1) were mutated based on the reported structure. An in vivo analysis of strains harboring these mutations revealed the structural determinants of the redox response during the light/dark transitions. In addition, we established a half day purification method for the entire CF(o)CF(1) complex from C. reinhardtii and subsequently examined ATP synthesis activity by the acid–base transition method. We found that truncation of the β-hairpin domain resulted in a loss of redox regulation of ATP synthesis (i.e., constitutively active state) despite retaining redox-sensitive Cys residues. In contrast, truncation of the redox loop domain containing the Cys residues resulted in a marked decrease in the activity. Based on this mutation analysis, we propose a model of redox regulation of the ATP synthesis reaction by the cooperative function of the β-hairpin and the redox loop domains specific to CF(o)CF(1). National Academy of Sciences 2023-01-30 2023-02-07 /pmc/articles/PMC9964038/ /pubmed/36716358 http://dx.doi.org/10.1073/pnas.2218187120 Text en Copyright © 2023 the Author(s). Published by PNAS. https://creativecommons.org/licenses/by-nc-nd/4.0/This article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND) (https://creativecommons.org/licenses/by-nc-nd/4.0/) . |
spellingShingle | Biological Sciences Akiyama, Kentaro Ozawa, Shin-Ichiro Takahashi, Yuichiro Yoshida, Keisuke Suzuki, Toshiharu Kondo, Kumiko Wakabayashi, Ken-ichi Hisabori, Toru Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title | Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title_full | Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title_fullStr | Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title_full_unstemmed | Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title_short | Two specific domains of the γ subunit of chloroplast F(o)F(1) provide redox regulation of the ATP synthesis through conformational changes |
title_sort | two specific domains of the γ subunit of chloroplast f(o)f(1) provide redox regulation of the atp synthesis through conformational changes |
topic | Biological Sciences |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9964038/ https://www.ncbi.nlm.nih.gov/pubmed/36716358 http://dx.doi.org/10.1073/pnas.2218187120 |
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