article epub_ahead yes Simon Mayer author Marianna Fanouria Mitsioni author Paul Robin author 48c58128-57b0-11ee-9095-dc28fd97fc1d0000-0002-5728-9189 Lukas Van Den Heuvel author Nathan Ronceray author Maria Jose Marcaida author Luciano A. Abriata author Lucien F. Krapp author Jana S. Anton author Sarah Soussou author Justin Jeanneret-Grosjean author Alessandro Fulciniti author Alexia Möller author Sarah Vacle author Lely Feletti author Henry Brinkerhoff author Andrew H. Laszlo author Jens H. Gundlach author Theo Emmerich author Matteo Dal Peraro author Aleksandra Radenovic author EdHa department β-Barrel nanopores are involved in crucial biological processes, from ATP export in mitochondria to bacterial resistance, and represent a promising platform for emerging sequencing technologies. However, in contrast to ion channels, the understanding of the fundamental principles governing ion transport through these nanopores remains largely unexplored. Here we integrate experimental, numerical and theoretical approaches to elucidate ion transport mechanisms in β-barrel nanopores. We identify and characterize two distinct nonlinear phenomena: open-pore rectification and gating. Through extensive mutation analysis of aerolysin nanopores, we demonstrate that open-pore rectification is caused by ionic accumulation driven by the distribution of lumen charges. In addition, we provide converging evidence suggesting that gating is controlled by electric fields dissociating counterions from lumen charges, promoting local structural deformations. Our findings establish a rigorous framework for characterizing and understanding ion transport processes in protein-based nanopores, enabling the design of adaptable nanofluidic biotechnologies. We illustrate this by optimizing an aerolysin mutant for computing applications. Springer Nature2025 eng 1748-3387 1748-3395 41219410 00161169890000110.1038/s41565-025-02052-6 S. Mayer <i>et al.</i>, “Lumen charge governs gated ion transport in β-barrel nanopores,” <i>Nature Nanotechnology</i>. Springer Nature, 2025. Mayer, S., Mitsioni, M. F., Robin, P., Van Den Heuvel, L., Ronceray, N., Marcaida, M. J., … Radenovic, A. (2025). Lumen charge governs gated ion transport in β-barrel nanopores. <i>Nature Nanotechnology</i>. Springer Nature. <a href="https://doi.org/10.1038/s41565-025-02052-6">https://doi.org/10.1038/s41565-025-02052-6</a> Mayer, Simon, et al. “Lumen Charge Governs Gated Ion Transport in β-Barrel Nanopores.” <i>Nature Nanotechnology</i>, Springer Nature, 2025, doi:<a href="https://doi.org/10.1038/s41565-025-02052-6">10.1038/s41565-025-02052-6</a>. Mayer S, Mitsioni MF, Robin P, et al. Lumen charge governs gated ion transport in β-barrel nanopores. <i>Nature Nanotechnology</i>. 2025. doi:<a href="https://doi.org/10.1038/s41565-025-02052-6">10.1038/s41565-025-02052-6</a> Mayer, Simon, Marianna Fanouria Mitsioni, Paul Robin, Lukas Van Den Heuvel, Nathan Ronceray, Maria Jose Marcaida, Luciano A. Abriata, et al. “Lumen Charge Governs Gated Ion Transport in β-Barrel Nanopores.” <i>Nature Nanotechnology</i>. Springer Nature, 2025. <a href="https://doi.org/10.1038/s41565-025-02052-6">https://doi.org/10.1038/s41565-025-02052-6</a>. Mayer S, Mitsioni MF, Robin P, Van Den Heuvel L, Ronceray N, Marcaida MJ, Abriata LA, Krapp LF, Anton JS, Soussou S, Jeanneret-Grosjean J, Fulciniti A, Möller A, Vacle S, Feletti L, Brinkerhoff H, Laszlo AH, Gundlach JH, Emmerich T, Dal Peraro M, Radenovic A. 2025. Lumen charge governs gated ion transport in β-barrel nanopores. Nature Nanotechnology. S. Mayer, M.F. Mitsioni, P. Robin, L. Van Den Heuvel, N. Ronceray, M.J. Marcaida, L.A. Abriata, L.F. Krapp, J.S. Anton, S. Soussou, J. Jeanneret-Grosjean, A. Fulciniti, A. Möller, S. Vacle, L. Feletti, H. Brinkerhoff, A.H. Laszlo, J.H. Gundlach, T. Emmerich, M. Dal Peraro, A. Radenovic, Nature Nanotechnology (2025). 206702025-11-23T23:01:40Z2026-06-18T18:25:33Z