Molecular and cellular dissection of the oxysterol-binding protein cycle through a fluorescent inhibitor
| Autor: | Meng-Chen Tsai, Sandy Desrat, Melody Subra, Lucile Fleuriot, Van Cuong Pham, Romain Gautier, Joël Polidori, Marc Litaudon, Tiphaine Péresse, David Kovacs, Jérôme Bignon, Fanny Roussi, Delphine Debayle, Joëlle Bigay, Bruno Antonny, Bruno Mesmin |
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| Přispěvatelé: | Institut de Chimie des Substances Naturelles (ICSN), Institut de Chimie du CNRS (INC)-Université Paris-Saclay-Centre National de la Recherche Scientifique (CNRS), Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Vietnam Academy of Science and Technology (VAST) |
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Receptors Steroid protein drug interaction ORPphilin Endoplasmic Reticulum Biochemistry Fluorescence 03 medical and health sciences chemistry.chemical_compound Membrane Biology lipid-transfer protein Organelle Stilbenes Schweinfurthin Golgi Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Phosphatidylinositol Molecular Biology OSBP Lipid Transport lipid transport 030102 biochemistry & molecular biology Chemistry oxysterol-binding protein (OSBP) cholesterol Biological Transport Cell Biology Lipids Cell biology phosphoinositide Protein Transport 030104 developmental biology Förster resonance energy transfer fluorescence resonance energy transfer (FRET) Oxysterol-binding protein Carrier Proteins Plant lipid transfer proteins Intracellular Protein Binding trans-Golgi Network |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2020, ⟨10.1074/jbc.RA119.012012⟩ J Biol Chem |
| ISSN: | 0021-9258 1083-351X |
| Popis: | International audience; ORPphilins are bioactive natural products that strongly and selectively inhibit the growth of some cancer cell lines and are proposed to target intracellular lipid-transfer proteins of the oxysterol-binding protein (OSBP) family. These conserved proteins exchange key lipids, such as cholesterol and phopsphatidylinositol-4-phosphate (PI(4)P), between organelle membranes. Among ORPphilins, molecules of the schweinfurthin family interfere with intracellular lipid distribution and metabolism, but their functioning at the molecular level is poorly understood. We report here that cell line sensitivity to schweinfurthin G (SWG) is inversely proportional to cellular OSBP levels. By taking advantage of the intrinsic fluorescence of SWG, we followed its fate in cell cultures and show that its incorporation at the trans-Golgi network depends on cellular abundance of OSBP. Using in vitro membrane reconstitution systems and cellular imaging approaches, we also report that SWG inhibits specifically the lipid transfer activity of OSBP. As a consequence, post-Golgi trafficking, membrane cholesterol levels, and PI(4)P turnover were affected. Finally, using intermolecular FRET analysis, we demonstrate that SWG directly binds to the lipid-binding cavity of OSBP. Collectively these results describe SWG as a specific and intrinsically fluorescent pharmacological tool for dissecting OSBP properties at the cellular and molecular levels. Our findings indicate that SWG binds OSBP with nanomolar affinity, that this binding is sensitive to the membrane environment, and that SWG inhibits the OSBP-catalyzed lipid exchange cycle. |
| Databáze: | OpenAIRE |
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