title: Quantification of Phosphatidylinositol Phosphate Species in Purified Membranes.
abstract: Phosphoinositide lipids (PIPs) are required for various processes during macroautophagy, such as phagophore formation and autophagosome-lysosome fusion. Hence, quantification of the seven PIP species in autophagosome membranes is an important tool to understand how these lipids govern the transition of autophagosomes into autolysosomes. Here, we describe microscopic and mass spectrometry methods which, although designed to quantify the different PIP species on purified lysosomes, can also be applied to analyze autophagosomal PIPs.
authors.affiliation: Cell Biology Institute, University of Bonn, Bonn, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Jeschke
authors.affiliation: Priority Research Area Infections, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Zehethofer
authors.affiliation: Priority Research Area Infections, Research Center Borstel, Leibniz-Center for Medicine and Biosciences, Borstel, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Schwudke
authors.affiliation: Cell Biology Institute, University of Bonn, Bonn, Germany. Electronic address: albert.haas@uni-bonn.de.
authors.forename: A
authors.initials: A
authors.lastname: Haas
mesh_terms: D000071182:Autophagosomes; D002462:Cell Membrane / Q000737:chemistry*; D056148:Chromatography, Reverse-Phase / Q000379:methods; D056827:Endosomal Sorting Complexes Required for Transport / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D018129:Phosphatidylinositol Phosphates / Q000032:analysis* / Q000378:metabolism*; D010750:Phosphoproteins / Q000235:genetics / Q000378:metabolism
chemical_list: D056827:Endosomal Sorting Complexes Required for Transport; D018129:Phosphatidylinositol Phosphates; D010750:Phosphoproteins; C096164:hepatocyte growth factor-regulated tyrosine kinase substrate
keywords: Autophagosome; Autophagosome–lysosome fusion; FYVE; Lipid analysis; Lipid-binding domain; Lysosome; Mass spectrometry; Phosphatidylinositol 3-phosphate; Phosphatidylinositol 4-phosphate; Phosphoinositide