Towards the structure of yeast and mammalian P4-ATPases: Poster at the 15th International Conference on Na,K-ATPase and Related Transport ATPases, Otsu, Japan, 2017

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Towards the structure of yeast and mammalian P4-ATPases : Poster at the 15th International Conference on Na,K-ATPase and Related Transport ATPases, Otsu, Japan, 2017. / Lyons, Joseph; Laban, Milena; Mikkelsen, Stine; Ulstrup, Jakob; Mogensen, Louise S; Montigny, Cédric; Ash, Miriam-Rose; Molday, Robert S.; Frauenfeld, Jens; Andersen, Jens Peter; Lenoir, Guillaume; Nissen, Poul.

2017.

Research output: Contribution to conferencePosterResearch

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@conference{8d966b4e69c84b81830c58403a1203be,
title = "Towards the structure of yeast and mammalian P4-ATPases: Poster at the 15th International Conference on Na,K-ATPase and Related Transport ATPases, Otsu, Japan, 2017",
abstract = "P4-ATPases are members of the P-type ATPase superfamily that drive the inward translocation (flipping) of lipids within the membrane. These lipid flippase largely function as binary complexes with an auxiliary protein from the CDC50 family. The bulk of our knowledge has derived genetic and biochemical studies of yeast and mammalian P4-ATPases, in particular the phosphatidylserine (PS) transporting Drs2p/Cdc50p (Saccharomyces cerevisiae) and bATP8A2/CDC50A (Bos taurus). However, questions surrounding the mechanism of lipid translocation remain. To address this deficit in knowledge and to provide a basis for the analysis of reported mutagenesis data, we aim to solve the first molecular structures of the PS transporting P4-ATPases using electron microscopy. To date, negative stain EM analysis, on detergent, amphipol and saposin-lipoprotein nanoparticle (Salipro) reconstituted of both Drs2p/CDC50p and bATP8A2/CDC50A, has yielded comparable low-resolution envelopes of these two transporters, highlighting the bulk architecture of the complex. Current efforts and progress on the functional characterization and cryo-EM analysis of both lipid transporters reconstituted in Salipro are described. References1. Azouaoui, H., et al. PLoS One, 2014, 9, e1121762. Coleman, J. et al, J. Biol. Chem., 2009, 47, 326703. Vestergaard, A. L. et al., PNAS, 2014, 111, 1334-434. Azouaoui, H., et al. J. Biol. Chem., 2017, 292, 79545. Frauenfeld, J, et al, Nat. Methods, 13, 345Figure 1. Negative stain analysis of Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro. a-b) Representative 2D class averages of both proteins c) low resolution negative stain envelope of Drs2p/Cdc50p in salipro. The Na-K ATPase is used to highlight the architecture of the lipid flippase in the lipo-protein nanoparticle. d) Comparison of the low-resolution envelopes for Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro.",
author = "Joseph Lyons and Milena Laban and Stine Mikkelsen and Jakob Ulstrup and Mogensen, {Louise S} and C{\'e}dric Montigny and Miriam-Rose Ash and Molday, {Robert S.} and Jens Frauenfeld and Andersen, {Jens Peter} and Guillaume Lenoir and Poul Nissen",
year = "2017",
month = sep,
day = "24",
language = "Dansk",

}

RIS

TY - CONF

T1 - Towards the structure of yeast and mammalian P4-ATPases

T2 - Poster at the 15th International Conference on Na,K-ATPase and Related Transport ATPases, Otsu, Japan, 2017

AU - Lyons, Joseph

AU - Laban, Milena

AU - Mikkelsen, Stine

AU - Ulstrup, Jakob

AU - Mogensen, Louise S

AU - Montigny, Cédric

AU - Ash, Miriam-Rose

AU - Molday, Robert S.

AU - Frauenfeld, Jens

AU - Andersen, Jens Peter

AU - Lenoir, Guillaume

AU - Nissen, Poul

PY - 2017/9/24

Y1 - 2017/9/24

N2 - P4-ATPases are members of the P-type ATPase superfamily that drive the inward translocation (flipping) of lipids within the membrane. These lipid flippase largely function as binary complexes with an auxiliary protein from the CDC50 family. The bulk of our knowledge has derived genetic and biochemical studies of yeast and mammalian P4-ATPases, in particular the phosphatidylserine (PS) transporting Drs2p/Cdc50p (Saccharomyces cerevisiae) and bATP8A2/CDC50A (Bos taurus). However, questions surrounding the mechanism of lipid translocation remain. To address this deficit in knowledge and to provide a basis for the analysis of reported mutagenesis data, we aim to solve the first molecular structures of the PS transporting P4-ATPases using electron microscopy. To date, negative stain EM analysis, on detergent, amphipol and saposin-lipoprotein nanoparticle (Salipro) reconstituted of both Drs2p/CDC50p and bATP8A2/CDC50A, has yielded comparable low-resolution envelopes of these two transporters, highlighting the bulk architecture of the complex. Current efforts and progress on the functional characterization and cryo-EM analysis of both lipid transporters reconstituted in Salipro are described. References1. Azouaoui, H., et al. PLoS One, 2014, 9, e1121762. Coleman, J. et al, J. Biol. Chem., 2009, 47, 326703. Vestergaard, A. L. et al., PNAS, 2014, 111, 1334-434. Azouaoui, H., et al. J. Biol. Chem., 2017, 292, 79545. Frauenfeld, J, et al, Nat. Methods, 13, 345Figure 1. Negative stain analysis of Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro. a-b) Representative 2D class averages of both proteins c) low resolution negative stain envelope of Drs2p/Cdc50p in salipro. The Na-K ATPase is used to highlight the architecture of the lipid flippase in the lipo-protein nanoparticle. d) Comparison of the low-resolution envelopes for Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro.

AB - P4-ATPases are members of the P-type ATPase superfamily that drive the inward translocation (flipping) of lipids within the membrane. These lipid flippase largely function as binary complexes with an auxiliary protein from the CDC50 family. The bulk of our knowledge has derived genetic and biochemical studies of yeast and mammalian P4-ATPases, in particular the phosphatidylserine (PS) transporting Drs2p/Cdc50p (Saccharomyces cerevisiae) and bATP8A2/CDC50A (Bos taurus). However, questions surrounding the mechanism of lipid translocation remain. To address this deficit in knowledge and to provide a basis for the analysis of reported mutagenesis data, we aim to solve the first molecular structures of the PS transporting P4-ATPases using electron microscopy. To date, negative stain EM analysis, on detergent, amphipol and saposin-lipoprotein nanoparticle (Salipro) reconstituted of both Drs2p/CDC50p and bATP8A2/CDC50A, has yielded comparable low-resolution envelopes of these two transporters, highlighting the bulk architecture of the complex. Current efforts and progress on the functional characterization and cryo-EM analysis of both lipid transporters reconstituted in Salipro are described. References1. Azouaoui, H., et al. PLoS One, 2014, 9, e1121762. Coleman, J. et al, J. Biol. Chem., 2009, 47, 326703. Vestergaard, A. L. et al., PNAS, 2014, 111, 1334-434. Azouaoui, H., et al. J. Biol. Chem., 2017, 292, 79545. Frauenfeld, J, et al, Nat. Methods, 13, 345Figure 1. Negative stain analysis of Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro. a-b) Representative 2D class averages of both proteins c) low resolution negative stain envelope of Drs2p/Cdc50p in salipro. The Na-K ATPase is used to highlight the architecture of the lipid flippase in the lipo-protein nanoparticle. d) Comparison of the low-resolution envelopes for Drs2p/Cdc50p and bATP8A2/CDC50A in Salipro.

M3 - Poster

ER -