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Structural basis for the modulation of MRP2 activity by phosphorylation and drugs

Author

Listed:
  • Tiziano Mazza

    (Imperial College London
    Research Complex at Harwell
    University of Calabria)

  • Theodoros I. Roumeliotis

    (The Institute of Cancer Research)

  • Elena Garitta

    (Queen Mary University of London)

  • David Drew

    (Stockholm University)

  • S. Tamir Rashid

    (Imperial College London)

  • Cesare Indiveri

    (University of Calabria
    CNR Institute of Biomembranes, Bioenergetics and Molecular Biotechnology (IBIOM))

  • Jyoti S. Choudhary

    (The Institute of Cancer Research)

  • Kenneth J. Linton

    (Queen Mary University of London)

  • Konstantinos Beis

    (Imperial College London
    Research Complex at Harwell)

Abstract

Multidrug resistance-associated protein 2 (MRP2/ABCC2) is a polyspecific efflux transporter of organic anions expressed in hepatocyte canalicular membranes. MRP2 dysfunction, in Dubin-Johnson syndrome or by off-target inhibition, for example by the uricosuric drug probenecid, elevates circulating bilirubin glucuronide and is a cause of jaundice. Here, we determine the cryo-EM structure of rat Mrp2 (rMrp2) in an autoinhibited state and in complex with probenecid. The autoinhibited state exhibits an unusual conformation for this class of transporter in which the regulatory domain is folded within the transmembrane domain cavity. In vitro phosphorylation, mass spectrometry and transport assays show that phosphorylation of the regulatory domain relieves this autoinhibition and enhances rMrp2 transport activity. The in vitro data is confirmed in human hepatocyte-like cells, in which inhibition of endogenous kinases also reduces human MRP2 transport activity. The drug-bound state reveals two probenecid binding sites that suggest a dynamic interplay with autoinhibition. Mapping of the Dubin-Johnson mutations onto the rodent structure indicates that many may interfere with the transition between conformational states.

Suggested Citation

  • Tiziano Mazza & Theodoros I. Roumeliotis & Elena Garitta & David Drew & S. Tamir Rashid & Cesare Indiveri & Jyoti S. Choudhary & Kenneth J. Linton & Konstantinos Beis, 2024. "Structural basis for the modulation of MRP2 activity by phosphorylation and drugs," Nature Communications, Nature, vol. 15(1), pages 1-14, December.
  • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-46392-8
    DOI: 10.1038/s41467-024-46392-8
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    References listed on IDEAS

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    1. Alicia Lundby & Anna Secher & Kasper Lage & Nikolai B. Nordsborg & Anatoliy Dmytriyev & Carsten Lundby & Jesper V. Olsen, 2012. "Quantitative maps of protein phosphorylation sites across 14 different rat organs and tissues," Nature Communications, Nature, vol. 3(1), pages 1-10, January.
    2. Nitesh Kumar Khandelwal & Cinthia R. Millan & Samantha I. Zangari & Samantha Avila & Dewight Williams & Tarjani M. Thaker & Thomas M. Tomasiak, 2022. "The structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation," Nature Communications, Nature, vol. 13(1), pages 1-11, December.
    3. Qin Yu & Dongchun Ni & Julia Kowal & Ioannis Manolaridis & Scott M. Jackson & Henning Stahlberg & Kaspar P. Locher, 2021. "Structures of ABCG2 under turnover conditions reveal a key step in the drug transport mechanism," Nature Communications, Nature, vol. 12(1), pages 1-12, December.
    4. Jasmeen Oberoi & Xavi Aran Guiu & Emily A. Outwin & Pascale Schellenberger & Theodoros I. Roumeliotis & Jyoti S. Choudhary & Laurence H. Pearl, 2022. "HSP90-CDC37-PP5 forms a structural platform for kinase dephosphorylation," Nature Communications, Nature, vol. 13(1), pages 1-13, December.
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