Cryo-EM structure of the entire FtsH-HflKC AAA protease complex
The membrane-bound AAA protease FtsH is the key player controlling protein quality in bacteria. Two single-pass membrane proteins, HflK and HflC, interact with FtsH to modulate its proteolytic activity. Here, we present structure of the entire FtsH-HflKC complex, comprising 12 copies of both HflK an...
| Main Authors: | , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2022
|
| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/160820 |
| _version_ | 1826120339773456384 |
|---|---|
| author | Qiao, Zhu Yokoyama, Tatsuhiko Yan, Xin-Fu Beh, Ing Tsyr Shi, Jian Basak, Sandip Akiyama, Yoshinori Gao, Yong-Gui |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Qiao, Zhu Yokoyama, Tatsuhiko Yan, Xin-Fu Beh, Ing Tsyr Shi, Jian Basak, Sandip Akiyama, Yoshinori Gao, Yong-Gui |
| author_sort | Qiao, Zhu |
| collection | NTU |
| description | The membrane-bound AAA protease FtsH is the key player controlling protein quality in bacteria. Two single-pass membrane proteins, HflK and HflC, interact with FtsH to modulate its proteolytic activity. Here, we present structure of the entire FtsH-HflKC complex, comprising 12 copies of both HflK and HflC, all of which interact reciprocally to form a cage, as well as four FtsH hexamers with periplasmic domains and transmembrane helices enclosed inside the cage and cytoplasmic domains situated at the base of the cage. FtsH K61/D62/S63 in the β2-β3 loop in the periplasmic domain directly interact with HflK, contributing to complex formation. Pull-down and in vivo enzymatic activity assays validate the importance of the interacting interface for FtsH-HflKC complex formation. Structural comparison with the substrate-bound human m-AAA protease AFG3L2 offers implications for the HflKC cage in modulating substrate access to FtsH. Together, our findings provide a better understanding of FtsH-type AAA protease holoenzyme assembly and regulation. |
| first_indexed | 2024-10-01T05:14:33Z |
| format | Journal Article |
| id | ntu-10356/160820 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T05:14:33Z |
| publishDate | 2022 |
| record_format | dspace |
| spelling | ntu-10356/1608202023-02-28T17:12:22Z Cryo-EM structure of the entire FtsH-HflKC AAA protease complex Qiao, Zhu Yokoyama, Tatsuhiko Yan, Xin-Fu Beh, Ing Tsyr Shi, Jian Basak, Sandip Akiyama, Yoshinori Gao, Yong-Gui School of Biological Sciences NTU Institute of Structural Biology Science::Biological sciences AAA Protease Protein Quality Control The membrane-bound AAA protease FtsH is the key player controlling protein quality in bacteria. Two single-pass membrane proteins, HflK and HflC, interact with FtsH to modulate its proteolytic activity. Here, we present structure of the entire FtsH-HflKC complex, comprising 12 copies of both HflK and HflC, all of which interact reciprocally to form a cage, as well as four FtsH hexamers with periplasmic domains and transmembrane helices enclosed inside the cage and cytoplasmic domains situated at the base of the cage. FtsH K61/D62/S63 in the β2-β3 loop in the periplasmic domain directly interact with HflK, contributing to complex formation. Pull-down and in vivo enzymatic activity assays validate the importance of the interacting interface for FtsH-HflKC complex formation. Structural comparison with the substrate-bound human m-AAA protease AFG3L2 offers implications for the HflKC cage in modulating substrate access to FtsH. Together, our findings provide a better understanding of FtsH-type AAA protease holoenzyme assembly and regulation. Ministry of Education (MOE) Published version This work was supported by a Tier II grant MOE2019-T2-2-099 and a Tier 1 grant RG108/20 from the Ministry of Education (MOE) of Singapore (Y.-G.G.). 2022-08-03T05:01:59Z 2022-08-03T05:01:59Z 2022 Journal Article Qiao, Z., Yokoyama, T., Yan, X., Beh, I. T., Shi, J., Basak, S., Akiyama, Y. & Gao, Y. (2022). Cryo-EM structure of the entire FtsH-HflKC AAA protease complex. Cell Reports, 39(9), 110890-. https://dx.doi.org/10.1016/j.celrep.2022.110890 2211-1247 https://hdl.handle.net/10356/160820 10.1016/j.celrep.2022.110890 35649372 2-s2.0-85131144891 9 39 110890 en MOE2019-T2-2-099 RG108/20 Cell Reports © 2022 The Author(s). This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). application/pdf |
| spellingShingle | Science::Biological sciences AAA Protease Protein Quality Control Qiao, Zhu Yokoyama, Tatsuhiko Yan, Xin-Fu Beh, Ing Tsyr Shi, Jian Basak, Sandip Akiyama, Yoshinori Gao, Yong-Gui Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title | Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title_full | Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title_fullStr | Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title_full_unstemmed | Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title_short | Cryo-EM structure of the entire FtsH-HflKC AAA protease complex |
| title_sort | cryo em structure of the entire ftsh hflkc aaa protease complex |
| topic | Science::Biological sciences AAA Protease Protein Quality Control |
| url | https://hdl.handle.net/10356/160820 |
| work_keys_str_mv | AT qiaozhu cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT yokoyamatatsuhiko cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT yanxinfu cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT behingtsyr cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT shijian cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT basaksandip cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT akiyamayoshinori cryoemstructureoftheentireftshhflkcaaaproteasecomplex AT gaoyonggui cryoemstructureoftheentireftshhflkcaaaproteasecomplex |