Bidirectional communication between nucleotide and substrate binding sites in a type IV multidrug ABC transporter
ATP-binding cassette (ABC) transporters use ATP to transport substrates across membranes. In type IV ABC transporters, which include many multidrug resistance (MDR) pumps, communication between nucleotide-binding domains (NBDs) and transmembrane domains (TMDs) is mediated via large intracellular domains containing 'coupling helices'. However, how ATP hydrolysis and substrate transport are functionally coordinated remains unclear. In the bacterial type IV MDR transporter BmrA, we identify a conserved residue cluster at the NBD/TMD interface centered on W413. Mutation of this tryptophan uncouples ATP hydrolysis from transport activity. Mutagenesis, functional assays, nuclear magnetic resonance spectroscopy, hydrogen-deuterium exchange mass spectrometry, and photo-induced electron-transfer fluorescence correlation spectroscopy show that the cluster forms a bidirectional communication hinge that relays signals between the NBD and TMD via coupling helix 2. Hinge mutations affect both local and global dynamics thereby influencing transporter activity. These findings uncover an allosteric pathway critical for functional coupling in multidomain ABC transporters.
Authors
- Victor Hugo Pérez Carrillo (ORCID: https://orcid.org/0000-0003-3580-4941)
- Cédric Orelle (ORCID: https://orcid.org/0000-0003-3418-3290)
- Margot Di Cesare (ORCID: https://orcid.org/0000-0002-7883-852X)
- Waqas Javed (ORCID: https://orcid.org/0000-0002-3381-4794)
- Hannes Neuweiler (ORCID: https://orcid.org/0000-0002-6657-001X)
- Ute A. Hellmich (ORCID: https://orcid.org/0000-0001-7162-285X)
- Dania Rose-Sperling (ORCID: https://orcid.org/0000-0002-2214-3243)
- Jean‐Michel Jault (ORCID: https://orcid.org/0000-0003-1743-2777)
- Julien Marcoux (ORCID: https://orcid.org/0000-0001-7321-7436)
Institutions
- Université Claude Bernard Lyon 1 (FR)
- Goethe University Frankfurt (DE)
- Centre National de la Recherche Scientifique (FR)
- University of Würzburg (DE)
- Structural and Molecular Basis of Infectious Systems (FR)
- Institut de Pharmacologie et de Biologie Structurale (FR)
- French Clinical Research Infrastructure Network (FR)
- Friedrich Schiller University Jena (DE)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-11-11
- DOI
- https://doi.org/10.1038/s41467-025-65037-y
- Citations
- 1
- Primary Topic
- Drug Transport and Resistance Mechanisms
- Type
- article
- Field-Weighted Citation Impact
- 1.05
Funders
- Friedrich-Schiller-Universität Jena
- European Commission
- Deutscher Akademischer Austauschdienst
- Deutsche Forschungsgemeinschaft
- Agence Nationale de la Recherche
- Institut National de la Santé et de la Recherche Médicale
- Fondation pour la Recherche Médicale
- Consejo Nacional de Ciencia y Tecnología
- Centre National de la Recherche Scientifique
- Hans Böckler Stiftung
- École Normale Supérieure de Lyon
- European Regional Development Fund