In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC
Abstract The MacAB-TolC tripartite efflux pump is widely distributed in Gram-negative bacteria, extruding antibiotics and virulence factors that lead to multidrug resistance and pathogenicity. However, the in situ structure and assembly mechanism of MacAB-TolC remain unclear. Here, we resolve the in...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-06-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-08236-z |
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| author | Tong Huo Wenfang Zhang Zhili Yu Wei Zheng Yaoming Wu Qiuyu Ren Zekai Wan Junli Cao Zhao Wang Xiaodong Shi |
| author_facet | Tong Huo Wenfang Zhang Zhili Yu Wei Zheng Yaoming Wu Qiuyu Ren Zekai Wan Junli Cao Zhao Wang Xiaodong Shi |
| author_sort | Tong Huo |
| collection | DOAJ |
| description | Abstract The MacAB-TolC tripartite efflux pump is widely distributed in Gram-negative bacteria, extruding antibiotics and virulence factors that lead to multidrug resistance and pathogenicity. However, the in situ structure and assembly mechanism of MacAB-TolC remain unclear. Here, we resolve the in situ structures of the MacAB-TolC efflux pump in Escherichia coli by electron cryo-tomography and subtomogram averaging. In the cells without antibiotic treatment, we observe a fully assembled MacAB-TolC pump. When Escherichia coli cells are treated with erythromycin, in addition to the tripartite pumps, we also discover the emergence of MacA-TolC subcomplexes without MacB, indicating flexible binding of MacB in the presence of an antibiotic substrate, which is further validated by in vivo photo-crosslinking results. Together, our data suggest the in situ assembly process of MacAB-TolC starts from the formation of MacA-TolC subcomplex, and provides insights into the design of efflux pump inhibitors. |
| format | Article |
| id | doaj-art-aeadff8d90864d3b8033c7df29f1a3af |
| institution | OA Journals |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-aeadff8d90864d3b8033c7df29f1a3af2025-08-20T02:30:43ZengNature PortfolioCommunications Biology2399-36422025-06-01811810.1038/s42003-025-08236-zIn situ structure and assembly of the ABC-type tripartite pump MacAB-TolCTong Huo0Wenfang Zhang1Zhili Yu2Wei Zheng3Yaoming Wu4Qiuyu Ren5Zekai Wan6Junli Cao7Zhao Wang8Xiaodong Shi9Verna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of MedicineJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityVerna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of MedicineJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityVerna and Marrs McLean Department of Biochemistry and Molecular Pharmacology, Baylor College of MedicineJiangsu Province Key Laboratory of Anesthesiology and Jiangsu Province Key Laboratory of Anesthesia and Analgesia Application, Xuzhou Medical UniversityAbstract The MacAB-TolC tripartite efflux pump is widely distributed in Gram-negative bacteria, extruding antibiotics and virulence factors that lead to multidrug resistance and pathogenicity. However, the in situ structure and assembly mechanism of MacAB-TolC remain unclear. Here, we resolve the in situ structures of the MacAB-TolC efflux pump in Escherichia coli by electron cryo-tomography and subtomogram averaging. In the cells without antibiotic treatment, we observe a fully assembled MacAB-TolC pump. When Escherichia coli cells are treated with erythromycin, in addition to the tripartite pumps, we also discover the emergence of MacA-TolC subcomplexes without MacB, indicating flexible binding of MacB in the presence of an antibiotic substrate, which is further validated by in vivo photo-crosslinking results. Together, our data suggest the in situ assembly process of MacAB-TolC starts from the formation of MacA-TolC subcomplex, and provides insights into the design of efflux pump inhibitors.https://doi.org/10.1038/s42003-025-08236-z |
| spellingShingle | Tong Huo Wenfang Zhang Zhili Yu Wei Zheng Yaoming Wu Qiuyu Ren Zekai Wan Junli Cao Zhao Wang Xiaodong Shi In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC Communications Biology |
| title | In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC |
| title_full | In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC |
| title_fullStr | In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC |
| title_full_unstemmed | In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC |
| title_short | In situ structure and assembly of the ABC-type tripartite pump MacAB-TolC |
| title_sort | in situ structure and assembly of the abc type tripartite pump macab tolc |
| url | https://doi.org/10.1038/s42003-025-08236-z |
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