In situ structure of the AcrAB-ToIC efflux pump at subnanometer resolution

The tripartite AcrAB-ToIC assembly, which spans both the inner and outer membranes in Gram-negative bacteria, is an efflux pump that contributes to multidrug resistance. Here, we present the in situ structure of full-length Escherichia coli AcrAB-ToIC determined at 7 angstrom resolution by electron cryo-tomography. The ToIC channel penetrates the outer membrane bilayer through to the outer leaflet and exhibits two different configurations that differ by a 60 degrees rotation relative to the AcrB position in the pump assembly. AcrA protomers interact directly with the inner membrane and with AcrB via an interface located in proximity to the AcrB ligand-binding pocket. Our structural analysis suggests that these AcrA-bridged interactions underlie an allosteric mechanism for transmitting drug-evoked signals from AcrB to the ToIC channel within the pump. Our study demonstrates the power of in situ electron cryo-tomography, which permits critical insights into the function of bacterial efflux pumps.

Automated summary

This study presents the in situ structure of full-length Escherichia coli AcrAB-ToIC obtained by electron cryo-tomography, revealing the configuration and mechanism of this transmembrane efflux pump. The study suggests that AcrA acts as a bridge between the inner membrane and AcrB, enabling the transmission of drug-induced signals from AcrB to the ToIC channel within the pump.