Autoinhibition and regulation by phosphoinositides of ATP8B1, a human lipid flippase associated with intrahepatic cholestatic disorders
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Autoinhibition and regulation by phosphoinositides of ATP8B1, a human lipid flippase associated with intrahepatic cholestatic disorders. / Dieudonné, Thibaud; Herrera, Sara Abad; Laursen, Michelle Juknaviciute; Lejeune, Maylis; Stock, Charlott; Slimani, Kahina; Jaxel, Christine; Lyons, Joseph A.; Montigny, Cédric; Pomorski, Thomas Günther; Nissen, Poul; Lenoir, Guillaume.
I: eLife, Bind 11, e75272, 2022.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - Autoinhibition and regulation by phosphoinositides of ATP8B1, a human lipid flippase associated with intrahepatic cholestatic disorders
AU - Dieudonné, Thibaud
AU - Herrera, Sara Abad
AU - Laursen, Michelle Juknaviciute
AU - Lejeune, Maylis
AU - Stock, Charlott
AU - Slimani, Kahina
AU - Jaxel, Christine
AU - Lyons, Joseph A.
AU - Montigny, Cédric
AU - Pomorski, Thomas Günther
AU - Nissen, Poul
AU - Lenoir, Guillaume
N1 - Publisher Copyright: © 2022, Dieudonné et al.
PY - 2022
Y1 - 2022
N2 - P4-ATPases flip lipids from the exoplasmic to the cytosolic leaflet, thus maintaining lipid asymmetry in eukaryotic cell membranes. Mutations in several human P4-ATPase genes are associated with severe diseases, for example in ATP8B1 causing progressive familial intrahepatic cholestasis, a rare inherited disorder progressing toward liver failure. ATP8B1 forms a binary complex with CDC50A and displays a broad specificity to glycerophospholipids, but regulatory mechanisms are unknown. Here, we report functional studies and the cryo-EM structure of the human lipid flippase ATP8B1-CDC50A at 3.1 Å resolution. We find that ATP8B1 is autoinhibited by its N- and C-terminal tails, which form extensive interactions with the catalytic sites and flexible domain interfaces. Consistently, ATP hydrolysis is unleashed by truncation of the C-terminus, but also requires phosphoinositides, most markedly phosphatidylinositol-3,4,5-phosphate (PI(3,4,5)P3), and removal of both N- and C-termini results in full activation. Restored inhibition of ATP8B1 truncation constructs with a synthetic peptide mimicking the C-terminal segment further suggests molecular communication between N- and C-termini in the autoinhibition and demonstrates that the regulatory mechanism can be interfered with by exogenous compounds. A recurring (G/A)(Y/F)AFS motif of the C-terminal segment suggests that this mechanism is employed widely across P4-ATPase lipid flippases in plasma membrane and endomembranes.
AB - P4-ATPases flip lipids from the exoplasmic to the cytosolic leaflet, thus maintaining lipid asymmetry in eukaryotic cell membranes. Mutations in several human P4-ATPase genes are associated with severe diseases, for example in ATP8B1 causing progressive familial intrahepatic cholestasis, a rare inherited disorder progressing toward liver failure. ATP8B1 forms a binary complex with CDC50A and displays a broad specificity to glycerophospholipids, but regulatory mechanisms are unknown. Here, we report functional studies and the cryo-EM structure of the human lipid flippase ATP8B1-CDC50A at 3.1 Å resolution. We find that ATP8B1 is autoinhibited by its N- and C-terminal tails, which form extensive interactions with the catalytic sites and flexible domain interfaces. Consistently, ATP hydrolysis is unleashed by truncation of the C-terminus, but also requires phosphoinositides, most markedly phosphatidylinositol-3,4,5-phosphate (PI(3,4,5)P3), and removal of both N- and C-termini results in full activation. Restored inhibition of ATP8B1 truncation constructs with a synthetic peptide mimicking the C-terminal segment further suggests molecular communication between N- and C-termini in the autoinhibition and demonstrates that the regulatory mechanism can be interfered with by exogenous compounds. A recurring (G/A)(Y/F)AFS motif of the C-terminal segment suggests that this mechanism is employed widely across P4-ATPase lipid flippases in plasma membrane and endomembranes.
KW - autoinhibition
KW - biochemistry
KW - chemical biology
KW - Cryo-EM
KW - lipid flippase
KW - molecular biophysics
KW - P4-ATPases
KW - phosphoinositides
KW - progressive familial intrahepatic cholestasis
KW - S. cerevisiae
KW - structural biology
U2 - 10.7554/eLife.75272
DO - 10.7554/eLife.75272
M3 - Journal article
C2 - 35416773
AN - SCOPUS:85129778691
VL - 11
JO - eLife
JF - eLife
SN - 2050-084X
M1 - e75272
ER -
ID: 307295523