Structural characterization of the EmrAB-TolC efflux complex from E. coli

Gram-negative bacteria export a large variety of antimicrobial compounds by forming two-membrane spanning tripartite multidrug efflux systems composed of an inner membrane transporter, an outer membrane channel and a periplasmic adaptor protein. Here we present the co-expression, purification and first electron microscopy insights of the Escherichia coli EmrAB-TolC tripartite Major Facilitator Superfamily (MSF) efflux system as a whole complex stabilized by Amphipol polymer. The structure reveals a 33 nm long complex delineated by the Amphipol belt at both extremities. Comparison of projection structures of EmrAB-TolC and AcrAB-TolC indicates that the outer membrane protein TolC linked to the periplasmic adaptor EmrA protein form an extended periplasmic canal. The overall length of EmrAB-TolC complex is similar to that of AcrAB-TolC with a probable tip-to-tip interaction between EmrA and TolC unveiling how the adaptor protein connects TolC and EmrB embedded in the inner membrane.

Automated summary

This study investigates the structure and function of the Gram-negative bacteria efflux system, revealing the collaboration of inner membrane transporter, outer membrane channel, and periplasmic adaptor protein in the EmrAB-TolC tripartite system. Through electron microscopy, a 33 nm long complex stabilized by Amphipol belt was observed, suggesting a potential tip-to-tip interaction between EmrA and TolC.