Activation and substrate specificity of the human P4-ATPase ATP8B1

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Activation and substrate specificity of the human P4-ATPase ATP8B1. / Dieudonné, Thibaud; Kümmerer, Felix; Laursen, Michelle Juknaviciute; Stock, Charlott; Flygaard, Rasmus Kock; Khalid, Syma; Lenoir, Guillaume; Lyons, Joseph A.; Lindorff-Larsen, Kresten; Nissen, Poul.

In: Nature Communications, Vol. 14, No. 1, 7492, 2023.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Dieudonné, T, Kümmerer, F, Laursen, MJ, Stock, C, Flygaard, RK, Khalid, S, Lenoir, G, Lyons, JA, Lindorff-Larsen, K & Nissen, P 2023, 'Activation and substrate specificity of the human P4-ATPase ATP8B1', Nature Communications, vol. 14, no. 1, 7492. https://doi.org/10.1038/s41467-023-42828-9

APA

Dieudonné, T., Kümmerer, F., Laursen, M. J., Stock, C., Flygaard, R. K., Khalid, S., Lenoir, G., Lyons, J. A., Lindorff-Larsen, K., & Nissen, P. (2023). Activation and substrate specificity of the human P4-ATPase ATP8B1. Nature Communications, 14(1), [7492]. https://doi.org/10.1038/s41467-023-42828-9

Vancouver

Dieudonné T, Kümmerer F, Laursen MJ, Stock C, Flygaard RK, Khalid S et al. Activation and substrate specificity of the human P4-ATPase ATP8B1. Nature Communications. 2023;14(1). 7492. https://doi.org/10.1038/s41467-023-42828-9

Author

Dieudonné, Thibaud ; Kümmerer, Felix ; Laursen, Michelle Juknaviciute ; Stock, Charlott ; Flygaard, Rasmus Kock ; Khalid, Syma ; Lenoir, Guillaume ; Lyons, Joseph A. ; Lindorff-Larsen, Kresten ; Nissen, Poul. / Activation and substrate specificity of the human P4-ATPase ATP8B1. In: Nature Communications. 2023 ; Vol. 14, No. 1.

Bibtex

@article{ea3e5696f2ec4fd984e5173e3b6d9235,

title = "Activation and substrate specificity of the human P4-ATPase ATP8B1",

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 {\AA} 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)P3 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.",

author = "Thibaud Dieudonn{\'e} and Felix K{\"u}mmerer and Laursen, {Michelle Juknaviciute} and Charlott Stock and Flygaard, {Rasmus Kock} and Syma Khalid and Guillaume Lenoir and Lyons, {Joseph A.} and Kresten Lindorff-Larsen and Poul Nissen",

note = "Publisher Copyright: {\textcopyright} 2023, The Author(s).",

year = "2023",

doi = "10.1038/s41467-023-42828-9",

language = "English",

volume = "14",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "nature publishing group",

number = "1",

}

RIS

TY - JOUR

T1 - Activation and substrate specificity of the human P4-ATPase ATP8B1

AU - Dieudonné, Thibaud

AU - Kümmerer, Felix

AU - Laursen, Michelle Juknaviciute

AU - Stock, Charlott

AU - Flygaard, Rasmus Kock

AU - Khalid, Syma

AU - Lenoir, Guillaume

AU - Lyons, Joseph A.

AU - Lindorff-Larsen, Kresten

AU - Nissen, Poul

PY - 2023

Y1 - 2023

N2 - 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)P3 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.

AB - 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)P3 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.

U2 - 10.1038/s41467-023-42828-9

DO - 10.1038/s41467-023-42828-9

M3 - Journal article

C2 - 37980352

AN - SCOPUS:85176936385

VL - 14

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 7492

ER -

ID: 374403582

Department of Biology