A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1
Abstract Bcs1, a homo-heptameric transmembrane AAA-ATPase, facilitates folded Rieske iron-sulfur protein translocation across the inner mitochondrial membrane. Structures in different nucleotide states (ATPγS, ADP, apo) provided conformational snapshots, but the kinetics and structural transitions o...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-10-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-023-41806-5 |
| _version_ | 1797558064820256768 |
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| author | Yangang Pan Jingyu Zhan Yining Jiang Di Xia Simon Scheuring |
| author_facet | Yangang Pan Jingyu Zhan Yining Jiang Di Xia Simon Scheuring |
| author_sort | Yangang Pan |
| collection | DOAJ |
| description | Abstract Bcs1, a homo-heptameric transmembrane AAA-ATPase, facilitates folded Rieske iron-sulfur protein translocation across the inner mitochondrial membrane. Structures in different nucleotide states (ATPγS, ADP, apo) provided conformational snapshots, but the kinetics and structural transitions of the ATPase cycle remain elusive. Here, using high-speed atomic force microscopy (HS-AFM) and line scanning (HS-AFM-LS), we characterized single-molecule Bcs1 ATPase cycling. While the ATP conformation had ~5600 ms lifetime, independent of the ATP-concentration, the ADP/apo conformation lifetime was ATP-concentration dependent and reached ~320 ms at saturating ATP-concentration, giving a maximum turnover rate of 0.17 s−1. Importantly, Bcs1 ATPase cycle conformational changes occurred in concert. Furthermore, we propose that the transport mechanism involves opening the IMS gate through energetically costly straightening of the transmembrane helices, potentially driving rapid gate resealing. Overall, our results establish a concerted ATPase cycle mechanism in Bcs1, distinct from other AAA-ATPases that use a hand-over-hand mechanism. |
| first_indexed | 2024-03-10T17:25:11Z |
| format | Article |
| id | doaj.art-3f96619f16e4496cb8b3a916852c0658 |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-03-10T17:25:11Z |
| publishDate | 2023-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-3f96619f16e4496cb8b3a916852c06582023-11-20T10:11:21ZengNature PortfolioNature Communications2041-17232023-10-0114111310.1038/s41467-023-41806-5A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1Yangang Pan0Jingyu Zhan1Yining Jiang2Di Xia3Simon Scheuring4Department of Anesthesiology, Weill Cornell Medical CollegeLaboratory of Cell Biology, National Cancer Institute, National Institutes of HealthDepartment of Anesthesiology, Weill Cornell Medical CollegeLaboratory of Cell Biology, National Cancer Institute, National Institutes of HealthDepartment of Anesthesiology, Weill Cornell Medical CollegeAbstract Bcs1, a homo-heptameric transmembrane AAA-ATPase, facilitates folded Rieske iron-sulfur protein translocation across the inner mitochondrial membrane. Structures in different nucleotide states (ATPγS, ADP, apo) provided conformational snapshots, but the kinetics and structural transitions of the ATPase cycle remain elusive. Here, using high-speed atomic force microscopy (HS-AFM) and line scanning (HS-AFM-LS), we characterized single-molecule Bcs1 ATPase cycling. While the ATP conformation had ~5600 ms lifetime, independent of the ATP-concentration, the ADP/apo conformation lifetime was ATP-concentration dependent and reached ~320 ms at saturating ATP-concentration, giving a maximum turnover rate of 0.17 s−1. Importantly, Bcs1 ATPase cycle conformational changes occurred in concert. Furthermore, we propose that the transport mechanism involves opening the IMS gate through energetically costly straightening of the transmembrane helices, potentially driving rapid gate resealing. Overall, our results establish a concerted ATPase cycle mechanism in Bcs1, distinct from other AAA-ATPases that use a hand-over-hand mechanism.https://doi.org/10.1038/s41467-023-41806-5 |
| spellingShingle | Yangang Pan Jingyu Zhan Yining Jiang Di Xia Simon Scheuring A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 Nature Communications |
| title | A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 |
| title_full | A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 |
| title_fullStr | A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 |
| title_full_unstemmed | A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 |
| title_short | A concerted ATPase cycle of the protein transporter AAA-ATPase Bcs1 |
| title_sort | concerted atpase cycle of the protein transporter aaa atpase bcs1 |
| url | https://doi.org/10.1038/s41467-023-41806-5 |
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