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TitleIn situ structure and assembly of the ABC-type tripartite pump MacAB-TolC.
Journal, issue, pagesCommun Biol, Vol. 8, Issue 1, Page 848, Year 2025
Publish dateJun 3, 2025
AuthorsTong Huo / Wenfang Zhang / Zhili Yu / Wei Zheng / Yaoming Wu / Qiuyu Ren / Zekai Wan / Junli Cao / Zhao Wang / Xiaodong Shi /
PubMed AbstractThe 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 ...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.
External linksCommun Biol / PubMed:40461577 / PubMed Central
MethodsEM (subtomogram averaging)
Resolution12.0 - 14.0 Å
Structure data

EMDB-44384: The in situ structure of MacAB-TolC in native state
Method: EM (subtomogram averaging) / Resolution: 14.0 Å

EMDB-44385: The in situ structure of MacAB-TolC in erythromycin-present state
Method: EM (subtomogram averaging) / Resolution: 12.0 Å

Source
  • Escherichia coli (E. coli)

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