Cryo-EM structures capture the transport cycle of the P4-ATPase flippase
In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. Here, we report the cryo-electron microscopy...
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| Published in: | Science (American Association for the Advancement of Science) Vol. 365; no. 6458; p. 1149 |
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| Main Authors: | , , , |
| Format: | Journal Article |
| Language: | English |
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13.09.2019
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| ISSN: | 1095-9203, 1095-9203 |
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| Abstract | In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. |
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| AbstractList | In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases.In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. In eukaryotic membranes, type IV P-type adenosine triphosphatases (P4-ATPases) mediate the translocation of phospholipids from the outer to the inner leaflet and maintain lipid asymmetry, which is critical for membrane trafficking and signaling pathways. Here, we report the cryo-electron microscopy structures of six distinct intermediates of the human ATP8A1-CDC50a heterocomplex at resolutions of 2.6 to 3.3 angstroms, elucidating the lipid translocation cycle of this P4-ATPase. ATP-dependent phosphorylation induces a large rotational movement of the actuator domain around the phosphorylation site in the phosphorylation domain, accompanied by lateral shifts of the first and second transmembrane helices, thereby allowing phosphatidylserine binding. The phospholipid head group passes through the hydrophilic cleft, while the acyl chain is exposed toward the lipid environment. These findings advance our understanding of the flippase mechanism and the disease-associated mutants of P4-ATPases. |
| Author | Nishizawa, Tomohiro Hiraizumi, Masahiro Yamashita, Keitaro Nureki, Osamu |
| Author_xml | – sequence: 1 givenname: Masahiro orcidid: 0000-0002-4340-2937 surname: Hiraizumi fullname: Hiraizumi, Masahiro organization: Discovery Technology Laboratories, Innovative Research Division, Mitsubishi Tanabe Pharma Corporation, 1000 Kamoshida, Aoba-ku, Yokohama, 227-0033, Japan – sequence: 2 givenname: Keitaro orcidid: 0000-0002-5442-7582 surname: Yamashita fullname: Yamashita, Keitaro organization: RIKEN SPring-8 Center, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan – sequence: 3 givenname: Tomohiro orcidid: 0000-0001-7463-8398 surname: Nishizawa fullname: Nishizawa, Tomohiro email: t-2438@bs.s.u-tokyo.ac.jp, nureki@bs.s.u-tokyo.ac.jp organization: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. t-2438@bs.s.u-tokyo.ac.jp nureki@bs.s.u-tokyo.ac.jp – sequence: 4 givenname: Osamu orcidid: 0000-0003-1813-7008 surname: Nureki fullname: Nureki, Osamu email: t-2438@bs.s.u-tokyo.ac.jp, nureki@bs.s.u-tokyo.ac.jp organization: Department of Biological Sciences, Graduate School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan. t-2438@bs.s.u-tokyo.ac.jp nureki@bs.s.u-tokyo.ac.jp |
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| SubjectTerms | Adenosine Triphosphatases - chemistry Adenosine Triphosphate - chemistry Amino Acid Sequence Binding Sites Cryoelectron Microscopy HEK293 Cells Humans Membrane Proteins - chemistry Models, Molecular Phosphatidylserines Phospholipid Transfer Proteins - chemistry Phospholipids - metabolism Phosphorylation Protein Binding Protein Interaction Domains and Motifs Protein Structure, Quaternary Sequence Alignment |
| Title | Cryo-EM structures capture the transport cycle of the P4-ATPase flippase |
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