TY  - JOUR
A1  - Ornik-Cha, Alina
A1  - Wilhelm, Julia
A1  - Kobylka, Jessica
A1  - Sjuts, Hanno
A1  - Vargiu, Attilio V.
A1  - Malloci, Giuliano
A1  - Reitz, Julian
A1  - Seyberth, Anja
A1  - Frangakis, Achilleas S.
A1  - Pos, Klaas Martinus
T1  - Structural and functional analysis of the promiscuous AcrB and AdeB efflux pumps suggests different drug binding mechanisms
T2  - Nature Communications
N2  - Upon antibiotic stress Gram-negative pathogens deploy resistance-nodulation-cell division-type tripartite efflux pumps. These include a H+/drug antiporter module that recognizes structurally diverse substances, including antibiotics. Here, we show the 3.5 Å structure of subunit AdeB from the Acinetobacter baumannii AdeABC efflux pump solved by single-particle cryo-electron microscopy. The AdeB trimer adopts mainly a resting state with all protomers in a conformation devoid of transport channels or antibiotic binding sites. However, 10% of the protomers adopt a state where three transport channels lead to the closed substrate (deep) binding pocket. A comparison between drug binding of AdeB and Escherichia coli AcrB is made via activity analysis of 20 AdeB variants, selected on basis of side chain interactions with antibiotics observed in the AcrB periplasmic domain X-ray co-structures with fusidic acid (2.3 Å), doxycycline (2.1 Å) and levofloxacin (2.7 Å). AdeABC, compared to AcrAB-TolC, confers higher resistance to E. coli towards polyaromatic compounds and lower resistance towards antibiotic compounds.
KW  - Antimicrobial resistance
KW  - Cryoelectron microscopy
KW  - Enzyme mechanisms
KW  - Permeation and transport
KW  - X-ray crystallography
Y1  - 2021
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/69788
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-697889
SN  - 2041-1723
N1  - Atomic coordinates and structure factors reported in this paper have been deposited in the Protein Data Bank under accession numbers 7B8P (AdeB-OOO), 7B8Q (AdeB-L*OO), 7B8R (AcrBper/DARPin in complex with Doxycycline), 7B8S (AcrBper/DARPin in complex with fusidic acid), 7B8T (AcrBper/DARPin in complex with Levofloxacin). Atomic coordinates that were used and support the findings of this study are available in the Protein Data Bank under accession numbers 5ENS, 7KGI, 7KGH, 7KGG, 7KGD, 4DX5, 4DX7, 5NC5, 6OWS, 6IIA, 5LQ3, 4MT1, 3K07, 3K0I. Source data for Supplementary Figs. 1, 3, 4, 10-13 and 15 are provided with this paper. Source data are provided with this paper.
N1  - The research leading to these results was conducted as part of the Translocation consortium (www.translocation.eu) and has received support from the Innovative Medicines Joint Undertaking under Grant Agreement no. 115525, resources which are composed of financial contribution from the European Union seventh framework program (FP7/2007-2013).
N1  - Open Access funding enabled and organized by Projekt DEAL.
VL  - 12
IS  - art. 6919
SP  - 1
EP  - 14
PB  - Nature Publishing Group UK
CY  - [London]
ER  -