De-novo macrocyclic peptides dissect energy coupling of a heterodimeric ABC transporter by multimode allosteric inhibition
- Goethe University Frankfurt, Germany
- The University of Tokyo, Japan
Abstract
ATP-binding cassette (ABC) transporters constitute the largest family of primary active transporters involved in a multitude of physiological processes and human diseases. Despite considerable efforts, it remains unclear how ABC transporters harness the chemical energy of ATP to drive substrate transport across cell membranes. Here, by random nonstandard peptide integrated discovery (RaPID), we leveraged combinatorial macrocyclic peptides that target a heterodimeric ABC transport complex and explore fundamental principles of the substrate translocation cycle. High-affinity peptidic macrocycles bind conformationally selective and display potent multimode inhibitory effects. The macrocycles block the transporter either before or after unidirectional substrate export along a single conformational switch induced by ATP binding. Our study reveals mechanistic principles of ATP binding, conformational switching, and energy transduction for substrate transport of ABC export systems. We highlight the potential of de novo macrocycles as effective inhibitors for membrane proteins implicated in multidrug resistance, providing avenues for the next-generation of pharmaceuticals.
Data availability
All data denerated or analyzed during this study are included in the manuscript and support files. A source data file has been provided for Figure 1C (Sequencing Data), Figure 2-6, Figure 2-supplement figure 1,and Figure 5-supplement figure 1.
Article and author information
Author details
Funding
Deutsche Forschungsgemeinschaft (TA 157/12-1)
- Robert Tampé
Deutsche Forschungsgemeinschaft (CRC 807/P16)
- Robert Tampé
Deutsche Forschungsgemeinschaft (CRC 807/24)
- Nina Morgner
European Research Council (798121)
- Robert Tampé
JSPS Grants-in-Aid for Research Fellowship (P15333)
- Richard Obexer
Japan Society for the Promotion of Science (JP20H05618)
- Hiroaki Suga
Japan Society for the Promotion of Science (JP20am0101090)
- Hiroaki Suga
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Reviewing Editor
- David Drew, Stockholm University, Sweden
Version history
- Received: February 21, 2021
- Accepted: April 29, 2021
- Accepted Manuscript published: April 30, 2021 (version 1)
- Version of Record published: May 12, 2021 (version 2)
Copyright
© 2021, Stefan 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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De-novo macrocyclic peptides dissect energy coupling of a heterodimeric ABC transporter by multimode allosteric inhibition
eLife 10:e67732.
https://doi.org/10.7554/eLife.67732
