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Cited by 40Lipid Membrane Structure and BehaviorATP Synthase and ATPases ResearchDrug Transport and Resistance Mechanisms

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Activation and substrate specificity of the human P4-ATPase ATP8B1

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Dieudonné, Thibaud, Kümmerer, Felix, Laursen, Michelle Juknaviciute, Stock, Charlott, Flygaard, Rasmus Kock, Khalid, Syma, Lenoir, Guillaume, Lyons, Joseph A., Lindorff-Larsen, Kresten, Nissen, Poul·2023·Nature Communications
DOI 10.1038/s41467-023-42828-9PMC PMC10657443

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Abstract

Asymmetric distribution of phospholipids in eukaryotic membranes is essential for cell integrity, signaling pathways, and vesicular trafficking. P4-ATPases, also known as flippases, participate in creating and maintaining this asymmetry through active transport of phospholipids from the exoplasmic to the cytosolic leaflet. Here, we present a total of nine cryo-electron microscopy structures of the human flippase ATP8B1-CDC50A complex at 2.4 to 3.1 Å overall resolution, along with functional and computational studies, addressing the autophosphorylation steps from ATP, substrate recognition and occlusion, as well as a phosphoinositide binding site. We find that the P4-ATPase transport site is occupied by water upon phosphorylation from ATP. Additionally, we identify two different autoinhibited states, a closed and an outward-open conformation. Furthermore, we identify and characterize the PI(3,4,5)P 3 binding site of ATP8B1 in an electropositive pocket between transmembrane segments 5, 7, 8, and 10. Our study also highlights the structural basis of a broad lipid specificity of ATP8B1 and adds phosphatidylinositol as a transport substrate for ATP8B1. We report a critical role of the sn-2 ester bond of glycerophospholipids in substrate recognition by ATP8B1 through conserved S403. These findings provide fundamental insights into ATP8B1 catalytic cycle and regulation, and substrate recognition in P4-ATPases.

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Metadata

Authors
Dieudonné, T., Kümmerer, F., Laursen, M. J., Stock, C., Flygaard, R. K., Khalid, S., Lenoir, G., Lyons, J. A., Lindorff-Larsen, K., & Nissen, P.
Affiliations
  • Centre National de la Recherche Scientifique (FR)
  • University of Copenhagen (DK)
  • Aarhus University (DK)
  • University of Oxford (GB)
Corresponding author
  • Dieudonné, Thibaud
  • Nissen, Poul
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2023-11-18
Volume
14
Issue
1
Pages
7492-7492
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-023-42828-9
PMC
PMC10657443
OpenAlex
W4388794978
Cited by
40
References
2
Citations by year
2026: 16 · 2025: 16 · 2024: 7 · 2023: 1
FWCI
3.55
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Research at the University of Copenhagen (University of Copenhagen) (publishedVersion, cc-by)
  • HAL (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • Aarhus Universitet
  • Agence Nationale de la Recherche (ANR- 10-INSB-05, ANR-10, ANR-14-CE09-0022, ANR-10-INBS-0005)
  • Lundbeckfonden (R248‐2016‐2518, R310‐2018‐3713, 155-2015-2666, R155-2015-2666, 2015-2666)
  • Centre National de la Recherche Scientifique
  • Novo Nordisk Fonden (NNF18OC0054875)
  • French Infrastructure for Integrated Structural Biology (ANR-10- INSB-05, INSB-05)
  • H. Lundbeck A/S
  • Engineering and Physical Sciences Research Council (EP/X035603, EP/V030779)
  • H2020 Marie Skłodowska-Curie Actions
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › ATP Synthase and ATPases Research
  • Health Sciences › Medicine › Oncology › Drug Transport and Resistance Mechanisms
Keywords
ATP8B1, P4-ATPase, flippase, phospholipid asymmetry, membrane lipid transport, CDC50A, autophosphorylation, substrate recognition, phosphatidylinositol 3,4,5-trisphosphate, phosphatidylinositol, cryo-EM structure
MeSH terms
Adenosine Triphosphatases, Adenosine Triphosphate, Cell Membrane, Humans, Phospholipids, Substrate Specificity, Cryoelectron Microscopy, Phospholipid Transfer Proteins

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