|Entry||Database: EMDB / ID: EMD-0533|
|Title||AcrAB-TolC open state - In situ structure and assembly of multidrug efflux pump AcrAB-TolC|
|Sample||E.coli cell overexpressing AcrAB-TolC treated with AcrB inhibitor|
|Biological species||Escherichia coli BL21(DE3) (bacteria)|
|Method||subtomogram averaging / cryo EM / Resolution: 20 Å|
|Authors||Chen M / Shi X / Ludtke SJ / Wang Z|
|Funding support|| United States, 2 items |
|Citation||Journal: Nat Commun / Year: 2019|
Title: In situ structure and assembly of the multidrug efflux pump AcrAB-TolC.
Authors: Xiaodong Shi / Muyuan Chen / Zhili Yu / James M Bell / Hans Wang / Isaac Forrester / Heather Villarreal / Joanita Jakana / Dijun Du / Ben F Luisi / Steven J Ludtke / Zhao Wang /
Abstract: Multidrug efflux pumps actively expel a wide range of toxic substrates from the cell and play a major role in intrinsic and acquired drug resistance. In Gram-negative bacteria, these pumps form ...Multidrug efflux pumps actively expel a wide range of toxic substrates from the cell and play a major role in intrinsic and acquired drug resistance. In Gram-negative bacteria, these pumps form tripartite assemblies that span the cell envelope. However, the in situ structure and assembly mechanism of multidrug efflux pumps remain unknown. Here we report the in situ structure of the Escherichia coli AcrAB-TolC multidrug efflux pump obtained by electron cryo-tomography and subtomogram averaging. The fully assembled efflux pump is observed in a closed state under conditions of antibiotic challenge and in an open state in the presence of AcrB inhibitor. We also observe intermediate AcrAB complexes without TolC and discover that AcrA contacts the peptidoglycan layer of the periplasm. Our data point to a sequential assembly process in living bacteria, beginning with formation of the AcrAB subcomplex and suggest domains to target with efflux pump inhibitors.
|Structure viewer||EM map: |
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|File||Download / File: emd_0533.map.gz / Format: CCP4 / Size: 15.6 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 2.78 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire E.coli cell overexpressing AcrAB-TolC treated with AcrB inhibitor
|Entire||Name: E.coli cell overexpressing AcrAB-TolC treated with AcrB inhibitor|
Number of components: 1
-Component #1: cellular-component, E.coli cell overexpressing AcrAB-TolC treated...
|Cellular-component||Name: E.coli cell overexpressing AcrAB-TolC treated with AcrB inhibitor|
Recombinant expression: No
|Source||Species: Escherichia coli BL21(DE3) (bacteria)|
|Specimen||Specimen state: Cell / Method: cryo EM|
|Sample solution||pH: 7|
|Vitrification||Cryogen name: ETHANE|
-Electron microscopy imaging
|Imaging||Microscope: JEOL 3200FSC|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 3 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
-Atomic model buiding
|Modeling #1||Refinement protocol: rigid body|
Input PDB model: 5V5S
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