AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex
Abstract Protein translocation across cellular membranes is an essential and nano-scale dynamic process. In the bacterial cytoplasmic membrane, the core proteins in this process are a membrane protein complex, SecYEG, corresponding to the eukaryotic Sec61 complex, and a cytoplasmic protein, SecA ATP...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-54875-x |
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| _version_ | 1841544447552126976 |
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| author | Yui Kanaoka Takaharu Mori Wataru Nagaike Seira Itaya Yuto Nonaka Hidetaka Kohga Takamitsu Haruyama Yasunori Sugano Ryoji Miyazaki Muneyoshi Ichikawa Takayuki Uchihashi Tomoya Tsukazaki |
| author_facet | Yui Kanaoka Takaharu Mori Wataru Nagaike Seira Itaya Yuto Nonaka Hidetaka Kohga Takamitsu Haruyama Yasunori Sugano Ryoji Miyazaki Muneyoshi Ichikawa Takayuki Uchihashi Tomoya Tsukazaki |
| author_sort | Yui Kanaoka |
| collection | DOAJ |
| description | Abstract Protein translocation across cellular membranes is an essential and nano-scale dynamic process. In the bacterial cytoplasmic membrane, the core proteins in this process are a membrane protein complex, SecYEG, corresponding to the eukaryotic Sec61 complex, and a cytoplasmic protein, SecA ATPase. Despite more than three decades of extensive research on Sec proteins, from genetic experiments to cutting-edge single-molecule analyses, no study has visually demonstrated protein translocation. Here, we visualize the translocation, via one unit of a SecYEG-SecA-embedded nanodisc, of an unfolded substrate protein by high-speed atomic force microscopy (HS-AFM). Additionally, the uniform unidirectional distribution of nanodiscs on a mica substrate enables the HS-AFM image data analysis, revealing dynamic structural changes in the polypeptide-crosslinking domain of SecA between wide-open and closed states depending on nucleotides. The nanodisc-AFM approach will allow us to execute detailed analyses of Sec proteins as well as visualize nano-scale events of other membrane proteins. |
| format | Article |
| id | doaj-art-8cb77387317745819bfef5d74107c849 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-8cb77387317745819bfef5d74107c8492025-01-12T12:31:41ZengNature PortfolioNature Communications2041-17232025-01-0116111210.1038/s41467-024-54875-xAFM observation of protein translocation mediated by one unit of SecYEG-SecA complexYui Kanaoka0Takaharu Mori1Wataru Nagaike2Seira Itaya3Yuto Nonaka4Hidetaka Kohga5Takamitsu Haruyama6Yasunori Sugano7Ryoji Miyazaki8Muneyoshi Ichikawa9Takayuki Uchihashi10Tomoya Tsukazaki11Department of Physics, Graduate School of Science, Nagoya UniversityDepartment of Chemistry, Faculty of Science, Tokyo University of ScienceDepartment of Physics, Graduate School of Science, Nagoya UniversityNara Institute of Science and TechnologyDepartment of Physics, Graduate School of Science, Nagoya UniversityNara Institute of Science and TechnologyNara Institute of Science and TechnologyNara Institute of Science and TechnologyNara Institute of Science and TechnologyState Key Laboratory of Genetic Engineering, Collaborative Innovation Center of Genetics and Development, Department of Biochemistry and Biophysics, School of Life Sciences, Fudan UniversityDepartment of Physics, Graduate School of Science, Nagoya UniversityNara Institute of Science and TechnologyAbstract Protein translocation across cellular membranes is an essential and nano-scale dynamic process. In the bacterial cytoplasmic membrane, the core proteins in this process are a membrane protein complex, SecYEG, corresponding to the eukaryotic Sec61 complex, and a cytoplasmic protein, SecA ATPase. Despite more than three decades of extensive research on Sec proteins, from genetic experiments to cutting-edge single-molecule analyses, no study has visually demonstrated protein translocation. Here, we visualize the translocation, via one unit of a SecYEG-SecA-embedded nanodisc, of an unfolded substrate protein by high-speed atomic force microscopy (HS-AFM). Additionally, the uniform unidirectional distribution of nanodiscs on a mica substrate enables the HS-AFM image data analysis, revealing dynamic structural changes in the polypeptide-crosslinking domain of SecA between wide-open and closed states depending on nucleotides. The nanodisc-AFM approach will allow us to execute detailed analyses of Sec proteins as well as visualize nano-scale events of other membrane proteins.https://doi.org/10.1038/s41467-024-54875-x |
| spellingShingle | Yui Kanaoka Takaharu Mori Wataru Nagaike Seira Itaya Yuto Nonaka Hidetaka Kohga Takamitsu Haruyama Yasunori Sugano Ryoji Miyazaki Muneyoshi Ichikawa Takayuki Uchihashi Tomoya Tsukazaki AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex Nature Communications |
| title | AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex |
| title_full | AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex |
| title_fullStr | AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex |
| title_full_unstemmed | AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex |
| title_short | AFM observation of protein translocation mediated by one unit of SecYEG-SecA complex |
| title_sort | afm observation of protein translocation mediated by one unit of secyeg seca complex |
| url | https://doi.org/10.1038/s41467-024-54875-x |
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