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A key residue of the extracellular gate provides quality control contributing to ABCG substrate specificity

Author

Listed:
  • Jian Xia

    (Department of Biology
    Martin Luther University Halle-Wittenberg)

  • Alexandra Siffert

    (Department of Biology
    University of Zürich)

  • Odalys Torres

    (Semmelweis University)

  • Francesca Iacobini

    (Department of Biology)

  • Joanna Banasiak

    (Polish Academy of Sciences)

  • Konrad Pakuła

    (Polish Academy of Sciences)

  • Jörg Ziegler

    (Leibniz Institute of Plant Biochemistry)

  • Sabine Rosahl

    (Leibniz Institute of Plant Biochemistry)

  • Noel Ferro

    (Friedrich-Vorwerk-Straße 13-15)

  • Michał Jasiński

    (Polish Academy of Sciences
    Poznan University of Life Sciences)

  • Tamás Hegedűs

    (Semmelweis University
    HUN-REN-SU)

  • Markus M. Geisler

    (Department of Biology
    HUN-REN-SU)

Abstract

For G-type ATP-binding cassette (ABC) transporters, a hydrophobic “di-leucine motif” as part of a hydrophobic extracellular gate has been described to separate a large substrate-binding cavity from a smaller upper cavity and proposed to act as a valve controlling drug extrusion. Here, we show that an L704F mutation in the hydrophobic extracellular gate of Arabidopsis ABCG36/PDR8/PEN3 uncouples the export of the auxin precursor indole-3-butyric acid (IBA) from that of the defense compound camalexin (CLX). Molecular dynamics simulations reveal increased free energy for CLX translocation in ABCG36L704F and reduced CLX contacts within the binding pocket proximal to the extracellular gate region. Mutation L704Y enables export of structurally related non-ABCG36 substrates, IAA, and indole, indicating allosteric communication between the extracellular gate and distant transport pathway regions. An evolutionary analysis identifies L704 as a Brassicaceae family-specific key residue of the extracellular gate that controls the identity of chemically similar substrates. In summary, our work supports the conclusion that L704 is a key residue of the extracellular gate that provides a final quality control contributing to ABCG substrate specificity, allowing for balance of growth-defense trade-offs.

Suggested Citation

  • Jian Xia & Alexandra Siffert & Odalys Torres & Francesca Iacobini & Joanna Banasiak & Konrad Pakuła & Jörg Ziegler & Sabine Rosahl & Noel Ferro & Michał Jasiński & Tamás Hegedűs & Markus M. Geisler, 2025. "A key residue of the extracellular gate provides quality control contributing to ABCG substrate specificity," Nature Communications, Nature, vol. 16(1), pages 1-16, December.
  • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-59518-3
    DOI: 10.1038/s41467-025-59518-3
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