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Cryo-EM of human P-glycoprotein reveals an intermediate occluded conformation during active drug transport

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  • Alan T. Culbertson

    (Harvard Medical School
    Iowa State University)

  • Maofu Liao

    (Harvard Medical School
    Southern University of Science and Technology
    Southern University of Science and Technology)

Abstract

P-glycoprotein (Pgp) is an important human multidrug transporter that contributes to pharmacokinetics and multidrug resistance. Despite decades of study, the conformation transition cycle of Pgp undergoing active drug transport is not defined, thus the precise relevance of all available Pgp structures to uninterrupted multidrug transport remains unclear. Here, we use cryo-EM of membrane-embedded human Pgp under continuous turnover conditions to analyze the conformational ensembles of Pgp transporting distinct substrates. These results delineate multiple conformations including inward-facing and closed conformations, highlighting the occluded conformation as a critical intermediate state between transporter closure and substrate release. A combination of structural, functional, and computational studies reveals the transmembrane helices 4 and 10 undergoing drastic rearrangement to coordinate substrate binding, occlusion, and release, and identifies a peripheral site involved in substrate capture and Pgp inhibition. Together, our results provide a set of snapshots of Pgp undergoing continuous drug transport, unveiling the intricate interplay between transporter dynamics and drug movement, and shed light on the mechanism of polyspecificity.

Suggested Citation

  • Alan T. Culbertson & Maofu Liao, 2025. "Cryo-EM of human P-glycoprotein reveals an intermediate occluded conformation during active drug transport," Nature Communications, Nature, vol. 16(1), pages 1-11, December.
  • Handle: RePEc:nat:natcom:v:16:y:2025:i:1:d:10.1038_s41467-025-58561-4
    DOI: 10.1038/s41467-025-58561-4
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    References listed on IDEAS

    as
    1. Wei Mi & Yanyan Li & Sung Hwan Yoon & Robert K. Ernst & Thomas Walz & Maofu Liao, 2017. "Structural basis of MsbA-mediated lipopolysaccharide transport," Nature, Nature, vol. 549(7671), pages 233-237, September.
    2. Atsushi Kodan & Tomohiro Yamaguchi & Toru Nakatsu & Keita Matsuoka & Yasuhisa Kimura & Kazumitsu Ueda & Hiroaki Kato, 2019. "Inward- and outward-facing X-ray crystal structures of homodimeric P-glycoprotein CmABCB1," Nature Communications, Nature, vol. 10(1), pages 1-12, December.
    3. Benjamin J. Orlando & Maofu Liao, 2020. "ABCG2 transports anticancer drugs via a closed-to-open switch," Nature Communications, Nature, vol. 11(1), pages 1-11, December.
    4. Mi Sun Jin & Michael L. Oldham & Qiuju Zhang & Jue Chen, 2012. "Crystal structure of the multidrug transporter P-glycoprotein from Caenorhabditis elegans," Nature, Nature, vol. 490(7421), pages 566-569, October.
    5. Stuti Sharma & Ruoyu Zhou & Li Wan & Shan Feng & KangKang Song & Chen Xu & Yanyan Li & Maofu Liao, 2021. "Mechanism of LolCDE as a molecular extruder of bacterial triacylated lipoproteins," Nature Communications, Nature, vol. 12(1), pages 1-11, December.
    6. Yanyan Li & Benjamin J. Orlando & Maofu Liao, 2019. "Structural basis of lipopolysaccharide extraction by the LptB2FGC complex," Nature, Nature, vol. 567(7749), pages 486-490, March.
    7. David Yin-wei Lin & Shuo Huang & Jue Chen, 2015. "Crystal structures of a polypeptide processing and secretion transporter," Nature, Nature, vol. 523(7561), pages 425-430, July.
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