The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis
The Na+translocating F1FO ATP synthase from Acetobacterium woodii shows a subunit stoichiometry of α3:β3:γ:δ:ε:a:b2:(c2/3)9:c1 and reveals an evolutionary path between synthases and pumps involving adaptations in the rotor c‐ring, which is composed of F‐ and vacuolar‐type c subunits in a stoichiomet...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
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2020
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| Online Access: | https://hdl.handle.net/10356/139309 |
| _version_ | 1811681228027330560 |
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| author | Bogdanović, Nebojša Trifunović, Dragan Sielaff, Hendrik Westphal, Lars Bhushan, Shashi Müller, Volker Grüber, Gerhard |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Bogdanović, Nebojša Trifunović, Dragan Sielaff, Hendrik Westphal, Lars Bhushan, Shashi Müller, Volker Grüber, Gerhard |
| author_sort | Bogdanović, Nebojša |
| collection | NTU |
| description | The Na+translocating F1FO ATP synthase from Acetobacterium woodii shows a subunit stoichiometry of α3:β3:γ:δ:ε:a:b2:(c2/3)9:c1 and reveals an evolutionary path between synthases and pumps involving adaptations in the rotor c‐ring, which is composed of F‐ and vacuolar‐type c subunits in a stoichiometry of 9 : 1. This hybrid turbine couples rotation with Na+ translocation in the FO part and rotation of the central stalk subunits γ‐ε to drive ATP synthesis in the catalytic α3:β3 headpiece. Here, we isolated a highly pure recombinant A. woodii F‐ATP synthase and present the first projected structure of this hybrid engine as determined by negative‐stain electron microscopy and single‐particle analysis. The uniqueness of the A. woodii F‐ATP synthase is also reflected by an extra 17 amino acid residues loop (195TSGKVKITEETKEEKSK211) in subunit γ. Deleting the loop‐encoding DNA sequence (γΔ195–211) and purifying the recombinant F‐ATP synthase γΔ195–211 mutant provided a platform to study its effect in enzyme stability and activity. The recombinant F‐ATP synthase γΔ195–211 mutant revealed the same subunit composition as the wild‐type enzyme and a minor reduction in ATP hydrolysis. When reconstituted into proteoliposomes ATP synthesis and Na+ transport were diminished, demonstrating the importance of the γ195–211 loop in both enzymatic processes. Based on a structural model, a coupling mechanism for this enzyme is proposed, highlighting the role of the γ‐loop. Finally, the γ195–211 loop of A. woodii is discussed in comparison with the extra γ‐loops of mycobacterial and chloroplasts F‐ATP synthases described to be involved in species‐specific regulatory mechanisms. |
| first_indexed | 2024-10-01T03:37:36Z |
| format | Journal Article |
| id | ntu-10356/139309 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:37:36Z |
| publishDate | 2020 |
| record_format | dspace |
| spelling | ntu-10356/1393092023-02-28T17:10:26Z The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis Bogdanović, Nebojša Trifunović, Dragan Sielaff, Hendrik Westphal, Lars Bhushan, Shashi Müller, Volker Grüber, Gerhard School of Biological Sciences NTU Institute of Structural Biology Science::Biological sciences::Biochemistry Science::Biological sciences::Biophysics Bioenergetics F-ATP Synthase The Na+translocating F1FO ATP synthase from Acetobacterium woodii shows a subunit stoichiometry of α3:β3:γ:δ:ε:a:b2:(c2/3)9:c1 and reveals an evolutionary path between synthases and pumps involving adaptations in the rotor c‐ring, which is composed of F‐ and vacuolar‐type c subunits in a stoichiometry of 9 : 1. This hybrid turbine couples rotation with Na+ translocation in the FO part and rotation of the central stalk subunits γ‐ε to drive ATP synthesis in the catalytic α3:β3 headpiece. Here, we isolated a highly pure recombinant A. woodii F‐ATP synthase and present the first projected structure of this hybrid engine as determined by negative‐stain electron microscopy and single‐particle analysis. The uniqueness of the A. woodii F‐ATP synthase is also reflected by an extra 17 amino acid residues loop (195TSGKVKITEETKEEKSK211) in subunit γ. Deleting the loop‐encoding DNA sequence (γΔ195–211) and purifying the recombinant F‐ATP synthase γΔ195–211 mutant provided a platform to study its effect in enzyme stability and activity. The recombinant F‐ATP synthase γΔ195–211 mutant revealed the same subunit composition as the wild‐type enzyme and a minor reduction in ATP hydrolysis. When reconstituted into proteoliposomes ATP synthesis and Na+ transport were diminished, demonstrating the importance of the γ195–211 loop in both enzymatic processes. Based on a structural model, a coupling mechanism for this enzyme is proposed, highlighting the role of the γ‐loop. Finally, the γ195–211 loop of A. woodii is discussed in comparison with the extra γ‐loops of mycobacterial and chloroplasts F‐ATP synthases described to be involved in species‐specific regulatory mechanisms. NRF (Natl Research Foundation, S’pore) Accepted version 2020-05-18T15:10:30Z 2020-05-18T15:10:30Z 2019 Journal Article Bogdanović, N., Trifunović, D., Sielaff, H., Westphal, L., Bhushan, S., Müller, V., & Grüber, G. (2019). The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis. FEBS Journal, 286(10), 1894-1907. doi:10.1111/febs.14793 1742-464X https://hdl.handle.net/10356/139309 10.1111/febs.14793 30791207 2-s2.0-85062782325 10 286 1894 1907 en FEBS Journal © 2019 Federation of European Biochemical Societies. All rights reserved. This is the accepted version of the following article: Bogdanović, N., Trifunović, D., Sielaff, H., Westphal, L., Bhushan, S., Müller, V., & Grüber, G. (2019). The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis. FEBS Journal, 286(10), 1894-1907. doi:10.1111/febs.14793, which has been published in final form at https://doi.org/10.1111/febs.14793 application/pdf |
| spellingShingle | Science::Biological sciences::Biochemistry Science::Biological sciences::Biophysics Bioenergetics F-ATP Synthase Bogdanović, Nebojša Trifunović, Dragan Sielaff, Hendrik Westphal, Lars Bhushan, Shashi Müller, Volker Grüber, Gerhard The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title | The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title_full | The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title_fullStr | The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title_full_unstemmed | The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title_short | The structural features of Acetobacterium woodii F‐ATP synthase reveal the importance of the unique subunit γ‐loop in Na+ translocation and ATP synthesis |
| title_sort | structural features of acetobacterium woodii f atp synthase reveal the importance of the unique subunit γ loop in na translocation and atp synthesis |
| topic | Science::Biological sciences::Biochemistry Science::Biological sciences::Biophysics Bioenergetics F-ATP Synthase |
| url | https://hdl.handle.net/10356/139309 |
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