title: Mass Assays to Quantify Bioactive PtdIns3P and PtdIns5P During Autophagic Responses.
abstract: Autophagy is a cellular process whereby cytoplasmic substrates are targeted for degradation in the lysosome via the membrane structures autophagosomes. This process is initiated by specific phosphoinositides, PtdIns3P and PtdIns5P, which play a key role in autophagy by recruiting effectors such as Atg18/WIPI2. Therefore, quantifying those lipids is important to better understand the assembly of the complex autophagic machinery. Herein, we describe in detail methods to quantify PtdIns3P and PtdIns5P by specific mass assays feasible in most laboratories.
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France.
authors.forename: J
authors.initials: J
authors.lastname: Viaud
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France.
authors.forename: G
authors.initials: G
authors.lastname: Chicanne
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France.
authors.forename: R
authors.initials: R
authors.lastname: Solinhac
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France.
authors.forename: K
authors.initials: K
authors.lastname: Hnia
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France.
authors.forename: F
authors.initials: F
authors.lastname: Gaits-Iacovoni
authors.affiliation: INSERM, U1048 and Université Toulouse 3, I2MC, Toulouse, France; CHU (Centre Hospitalier Universitaire) de Toulouse, Laboratoire d'Hématologie, Toulouse, France. Electronic address: bernard.payrastre@inserm.fr.
authors.forename: B
authors.initials: B
authors.lastname: Payrastre
mesh_terms: D000818:Animals; D001343:Autophagy / Q000502:physiology; D001345:Autoradiography / Q000379:methods; D008055:Lipids / Q000302:isolation & purification; D008967:Molecular Biology / Q000379:methods*; D018129:Phosphatidylinositol Phosphates / Q000032:analysis* / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D008055:Lipids; D018129:Phosphatidylinositol Phosphates; D011994:Recombinant Proteins; C055525:phosphatidylinositol 3-phosphate; C113567:phosphatidylinositol 5-phosphate; C514871:PIP4K2A protein, human; D017853:Phosphotransferases (Alcohol Group Acceptor)
keywords: Autophagy; Mass assay; Phosphoinositides; Phospholipids; PtdIns3P; PtdIns5P