Purification of AcrAB-TolC Multidrug Efflux Pump for Cryo-EM Analysis
The cell envelope of Gram-negative bacteria comprises an outer membrane, a cytoplasmic inner membrane, and an interstitial space. The tripartite multidrug transporter AcrAB-TolC, which uses proton electrochemical gradients to vectorially drive the efflux of drugs from the cell, spans this envelope. We describe here details of the methods used to prepare the recombinant tripartite assembly for high-resolution structure determination by cryo-EM.
Key wordsTransmembrane protein assembly Outer membrane protein Inner membrane Cryo-EM Three-dimensional structure Transport mechanism RND family
Wellcome Trust and Human Frontier Science Program provided funding for this work. The work was also supported by NIH grant NIH 5P41GM103832.