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ATP-dependent substrate transport by the ABC transporter MsbA is proton-coupled.

Published version
Peer-reviewed

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Authors

Singh, Himansha 
Velamakanni, Saroj 
Deery, Michael J 
Howard, Julie 
Wei, Shen L 

Abstract

ATP-binding cassette transporters mediate the transbilayer movement of a vast number of substrates in or out of cells in organisms ranging from bacteria to humans. Current alternating access models for ABC exporters including the multidrug and Lipid A transporter MsbA from Escherichia coli suggest a role for nucleotide as the fundamental source of free energy. These models involve cycling between conformations with inward- and outward-facing substrate-binding sites in response to engagement and hydrolysis of ATP at the nucleotide-binding domains. Here we report that MsbA also utilizes another major energy currency in the cell by coupling substrate transport to a transmembrane electrochemical proton gradient. The dependence of ATP-dependent transport on proton coupling, and the stimulation of MsbA-ATPase by the chemical proton gradient highlight the functional integration of both forms of metabolic energy. These findings introduce ion coupling as a new parameter in the mechanism of this homodimeric ABC transporter.

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Keywords

ATP-Binding Cassette Transporters, Adenosine Triphosphate, Amino Acid Sequence, Bacterial Proteins, Biological Transport, Chloramphenicol, Electrochemistry, Ethidium, Hydrolysis, Ions, Proteolipids, Protons, Substrate Specificity

Journal Title

Nat Commun

Conference Name

Journal ISSN

2041-1723
2041-1723

Volume Title

7

Publisher

Springer Science and Business Media LLC
Sponsorship
Biotechnology and Biological Sciences Research Council (BB/I002383/1)
Medical Research Council (G0401165)
Human Frontier Science Program (HFSP) (RPG0034/2013)
MRC (MC_PC_14116 v2)
Biotechnology and Biological Sciences Research Council (BB/K017713/1)
Himansha Singh is supported by the Cambridge Commonwealth, European and International Trust. Saroj Velamakanni was a recipient of a Cambridge Nehru Scholarship. Shen L. Wei was funded by the Cambridge Overseas Trust. This research in the Van Veen group was supported by Biotechnology and Biological Sciences Research Council (BBSRC) grant BB/I002383/1 and BB/C004663/1, Medical Research Council (MRC) grant G0401165 and by further support from the Human Frontier Science Program (HFSP) and the British Society for Antimicrobial Chemotherapy (BSAC).