The extracellular gate shapes the energy profile of an ABC exporter

Cedric A. J. Hutter, M. Hadi Timachi, Lea M. Hürlimann, Iwan Zimmermann, Pascal Egloff, Hendrik Göddeke, Svetlana Kucher, Saša Štefanić, Mikko Karttunen, Lars V. Schäfer, Enrica Bordignon () and Markus A. Seeger ()
Additional contact information
Cedric A. J. Hutter: University of Zurich
M. Hadi Timachi: Ruhr University Bochum
Lea M. Hürlimann: University of Zurich
Iwan Zimmermann: University of Zurich
Pascal Egloff: University of Zurich
Hendrik Göddeke: Ruhr University Bochum
Svetlana Kucher: Ruhr University Bochum
Saša Štefanić: University of Zurich
Mikko Karttunen: The University of Western Ontario
Lars V. Schäfer: Ruhr University Bochum
Enrica Bordignon: Ruhr University Bochum
Markus A. Seeger: University of Zurich

Nature Communications, 2019, vol. 10, issue 1, 1-13

Abstract: Abstract ABC exporters harness the energy of ATP to pump substrates across membranes. Extracellular gate opening and closure are key steps of the transport cycle, but the underlying mechanism is poorly understood. Here, we generated a synthetic single domain antibody (sybody) that recognizes the heterodimeric ABC exporter TM287/288 exclusively in the presence of ATP, which was essential to solve a 3.2 Å crystal structure of the outward-facing transporter. The sybody binds to an extracellular wing and strongly inhibits ATPase activity by shifting the transporter’s conformational equilibrium towards the outward-facing state, as shown by double electron-electron resonance (DEER). Mutations that facilitate extracellular gate opening result in a comparable equilibrium shift and strongly reduce ATPase activity and drug transport. Using the sybody as conformational probe, we demonstrate that efficient extracellular gate closure is required to dissociate the NBD dimer after ATP hydrolysis to reset the transporter back to its inward-facing state.

Date: 2019
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41467-019-09892-6 Abstract (text/html)

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-09892-6

Ordering information: This journal article can be ordered from
https://www.nature.com/ncomms/

DOI: 10.1038/s41467-019-09892-6

Access Statistics for this article

Nature Communications is currently edited by Nathalie Le Bot, Enda Bergin and Fiona Gillespie

More articles in Nature Communications from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().