Positively selected modifications in the pore of TbAQP2 allow pentamidine to enter Trypanosoma brucei
- University of Glasgow, United Kingdom
- University of Dundee, United Kingdom
- IOTA Pharmaceuticals Ltd, United Kingdom
- University of Liverpool, United Kingdom
- Georgia State University, United States
- University of Ghent, Belgium
- University of Cambridge, United Kingdom
- CSIC, Spain
- University of North Carolina at Chapel Hill, United States
Abstract
Mutations in the Trypanosoma brucei aquaporin AQP2 are associated with resistance to pentamidine and melarsoprol. We show that TbAQP2 but not TbAQP3 was positively selected for increased pore size from a common ancestor aquaporin. We demonstrate that TbAQP2's unique architecture permits pentamidine permeation through its central pore and show how specific mutations in highly conserved motifs affect drug permeation. Introduction of key TbAQP2 amino acids into TbAQP3 renders the latter permeable to pentamidine. Molecular dynamics demonstrates that permeation by dicationic pentamidine is energetically favourable in TbAQP2, driven by the membrane potential, although aquaporins are normally strictly impermeable for ionic species. We also identify the structural determinants that make pentamidine a permeant although most other diamidine drugs are excluded. Our results have wide-ranging implications for optimising antitrypanosomal drugs and averting cross-resistance. Moreover, these new insights in aquaporin permeation may allow the pharmacological exploitation of other members of this ubiquitous gene family.
Data availability
All data generated or analysed during this study are included in the manuscript and supporting files. Source data files have been provided that cover all figures and give raw data, averages, statistics etc.
Article and author information
Author details
Gustavo Daniel Campagnaro
Funding
Medical Research Council (G0701258)
- Harry P De Koning
National Institutes of Health (GM111749)
- David W Boykin
Medical Research Council (MR/R015791/1)
- Harry P De Koning
Scottish Universities Physics Alliance
- Ulrich Zachariae
Albaha University, Saudi Arabia
- Ali Alghamdi
Science Without Borders, Brazil (206385/2014-5)
- Gustavo Daniel Campagnaro
Wellcome (204697/Z/16/Z)
- Mark C Field
Medical Research Council
- Teresa Sprenger
Wellcome Trust and Royal Society (Sir Henry Dale fellowship)
- Simone Weyand
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- Christine Clayton, DKFZ-ZMBH Alliance, Germany
Publication history
- Received: February 26, 2020
- Accepted: August 6, 2020
- Accepted Manuscript published: August 7, 2020 (version 1)
- Accepted Manuscript updated: August 11, 2020 (version 2)
- Version of Record published: September 4, 2020 (version 3)
Copyright
© 2020, Alghamdi et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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Positively selected modifications in the pore of TbAQP2 allow pentamidine to enter Trypanosoma brucei
eLife 9:e56416.
https://doi.org/10.7554/eLife.56416
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