title: Method to Measure Sphingomyelin Synthase Activity Changes in Response to CD95L.
abstract: Sphingomyelin synthases 1 and 2 convert the anti-oncometabolite ceramide to sphingomyelin, the most abundant sphingolipid in plasma membrane. CD95L-induced ceramide increase is associated with the caspase-dependent inhibition of sphingomyelin synthesis, which enhances the mitochondrial route to apoptosis. Knocking down sphingomyelin synthase 1 or inhibiting sphingomyelin synthesis facilitates ceramide accumulation, cytochrome c release from mitochondria, and caspase-9 activation in cancer cell upon CD95L treatment. Here, we describe a method to monitor in situ sphingomyelin synthase activity changes triggered by CD95L.
authors.affiliation: INSERM UMR 1037, CRCT, 31037, Toulouse, France.|Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France.|Ecole Doctorale de Sciences et Technologies, Université Libanaise, Campus Universitaire de Rafic Hariri, Hadath, Lebanon.
authors.forename: Fatima
authors.initials: F
authors.lastname: Bilal
authors.affiliation: INSERM UMR 1037, CRCT, 31037, Toulouse, France.|Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France.|Université Toulouse III-Paul Sabatier, 118 route de Narbonne, 31062, Toulouse, France.
authors.forename: Michaël
authors.initials: M
authors.lastname: Pérès
authors.affiliation: INSERM UMR 1037, CRCT, 31037, Toulouse, France.|Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Andrieu-Abadie
authors.affiliation: INSERM UMR 1037, CRCT, 31037, Toulouse, France.|Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France.|Université Toulouse III-Paul Sabatier, 118 route de Narbonne, 31062, Toulouse, France.|Laboratoire de Biochimie, Institut Fédératif de Biologie, CHU Purpan, 31059, Toulouse, France.
authors.forename: Thierry
authors.initials: T
authors.lastname: Levade
authors.affiliation: Ecole Doctorale de Sciences et Technologies, Université Libanaise, Campus Universitaire de Rafic Hariri, Hadath, Lebanon.
authors.forename: Bassam
authors.initials: B
authors.lastname: Badran
authors.affiliation: Ecole Doctorale de Sciences et Technologies, Université Libanaise, Campus Universitaire de Rafic Hariri, Hadath, Lebanon.
authors.forename: Ahmad
authors.initials: A
authors.lastname: Daher
authors.affiliation: INSERM UMR 1037, CRCT, 31037, Toulouse, France. bruno.segui@inserm.fr.|Equipe Labellisée Ligue Contre Le Cancer, 31037, Toulouse, France. bruno.segui@inserm.fr.|Université Toulouse III-Paul Sabatier, 118 route de Narbonne, 31062, Toulouse, France. bruno.segui@inserm.fr.
authors.forename: Bruno
authors.initials: B
authors.lastname: Ségui
mesh_terms: D000818:Animals; D017209:Apoptosis; D002460:Cell Line; D002855:Chromatography, Thin Layer; D004789:Enzyme Activation; D057075:Enzyme Assays / Q000379:methods*; D053222:Fas Ligand Protein / Q000378:metabolism*; D051379:Mice; D011485:Protein Binding; D013109:Sphingomyelins / Q000302:isolation & purification / Q000378:metabolism; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism*
chemical_list: D053222:Fas Ligand Protein; D013109:Sphingomyelins; D017855:Transferases (Other Substituted Phosphate Groups); C070186:phosphatidylcholine-ceramide phosphocholine transferase
keywords: Apoptosis; Ceramide; In situ sphingomyelin synthase activity; Sphingomyelin