A High-Yield Co-Expression System for the Purification of an Intact Drs2p-Cdc50p Lipid Flippase Complex, Critically Dependent on and Stabilized by Phosphatidylinositol-4-Phosphate

Hassina Azouaoui; Cédric Montigny; Miriam-Rose Ash; Frank Fijalkowski; Aurore Jacquot; Christina Grønberg; Rosa L. López-Marqués; Michael G. Palmgren; Manuel Garrigos; Marc le Maire; Paulette Decottignies; Pontus Gourdon; Poul Nissen; Philippe Champeil; Guillaume Lenoir

doi:10.1371/journal.pone.0112176

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Research Article

A High-Yield Co-Expression System for the Purification of an Intact Drs2p-Cdc50p Lipid Flippase Complex, Critically Dependent on and Stabilized by Phosphatidylinositol-4-Phosphate

Hassina Azouaoui,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Cédric Montigny,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Miriam-Rose Ash,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
⨯
Frank Fijalkowski,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
⨯
Aurore Jacquot,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Christina Grønberg,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
⨯
Rosa L. López-Marqués,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark
⨯
Michael G. Palmgren,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark
⨯
Manuel Garrigos,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Marc le Maire,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Paulette Decottignies,

Affiliations CNRS, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619, Orsay, France, Univ Paris-Sud, Orsay, France
⨯
Pontus Gourdon,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
⨯
Poul Nissen,

Affiliations Centre for Membrane Pumps in Cells and Disease – PUMPKIN, Danish National Research Foundation, Aarhus, Denmark, Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark
⨯
Philippe Champeil,

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯
Guillaume Lenoir

* E-mail: guillaume.lenoir@cea.fr

Affiliations Univ Paris-Sud, UMR 8221, Orsay, France, CEA, iBiTec-S (Institut de Biologie et de Technologies de Saclay), SB2SM (Service de Bioénergétique, Biologie Structurale et Mécanismes), Laboratoire des Protéines Membranaires, Gif-sur-Yvette, France, CNRS, UMR 8221, Gif-sur-Yvette, France
⨯

A High-Yield Co-Expression System for the Purification of an Intact Drs2p-Cdc50p Lipid Flippase Complex, Critically Dependent on and Stabilized by Phosphatidylinositol-4-Phosphate

Hassina Azouaoui,
Cédric Montigny,
Miriam-Rose Ash,
Frank Fijalkowski,
Aurore Jacquot,
Christina Grønberg,
Rosa L. López-Marqués,
Michael G. Palmgren,
Manuel Garrigos,
Marc le Maire

Published: November 13, 2014
https://doi.org/10.1371/journal.pone.0112176

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