Titlebook: Intracellular Lipid Transport; Methods and Protocol Guillaume Drin Book 2019 Springer Science+Business Media, LLC, part of Springer Nature

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Determining the Lipid-Binding Specificity of SMP Domains: An ERMES Subunit as a Case Study,. The yeast Endoplasmic Reticulum–Mitochondrion Encounter Structure (ERMES) complex is composed of five subunits that tether the ER and mitochondria. Three ERMES subunits (i.e., Mdm12, Mmm1, and Mdm34) contain the synaptotagmin-like mitochondrial lipid-binding protein (SMP) domain. The SMP domain be

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In Vitro Measurement of Sphingolipid Intermembrane Transport Illustrated by GLTP Superfamily Membertracking of the lipid. Typically, direct tracking involves lipid labeling via attachment of fluorophores or introduction of radioactivity. Members of the GlycoLipid Transfer Protein (GLTP) superfamily are used to illustrate two broadly applicable methods for direct lipid tracking. One method relies

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In Vitro Strategy to Measure Sterol/Phosphatidylinositol-4-Phosphate Exchange Between Membranes,ily of lipid transfer proteins (LTPs), conserved among eukaryotes, are not all mere sterol transporters or sensors. Indeed, some of them are able to exchange sterol for phosphatidylinositol-4-phosphate (PI4P) or phosphatidylserine (PS) for PI4P between membranes and thereby to use PI4P metabolism to

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Monitoring Non-vesicular Transport of Phosphatidylserine and Phosphatidylinositol 4-Phosphate in Inbly tethered to the ER and also dynamically interacts with PM PI4P/PI(4,5)P. via its pleckstrin homology (PH) domain. We show that this method is able to detect PI4P and PS changes in the PM upon acute recruitment of an ORP5 construct to the PM. This method is convenient and robust, utilizing popula

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Intracellular and Plasma Membrane Cholesterol Labeling and Quantification Using Filipin and GFP-D4,. Second, to optically distinguish and quantify intracellular cholesterol accumulation, we have adapted the classical filipin cholesterol staining protocol. Indeed, we observed that treatment of living cells with methyl-β-cyclodextrin, a chemical known to extract cholesterol from the plasma membrane

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Determining the Lipid-Binding Specificity of SMP Domains: An ERMES Subunit as a Case Study,using lipid displacement and liposome flotation in vitro assays with liposomes as model biological membranes. These methods can be applied to the study and characterization of novel lipid-binding and lipid-transfer proteins.