title: Lipidomic profile of GM95 cell death induced by Clostridium perfringens alpha-toxin.
abstract: Clostridium perfringens alpha-toxin (ATX) is considered as a prototype of cytotoxic bacterial phospholipases C, and is the major virulence factor in C. perfringens-induced gas gangrene. It is known that, depending on the dose, ATX causes membrane disruption and cytolysis or only limited hydrolysis of its substrates. In the latter case, toxin activity leads to the unregulated generation of bioactive lipids that can ultimately induce cell death. We have characterized apoptosis and necrosis in highly ATX-sensitive, ganglioside-deficient cells exposed to different concentrations of ATX and we have studied the lipidomic profile of cells treated with ATX as compared to native cells to detect the main changes in the lipidomic profile and the possible involvement of lipid signals in cell death. ATX causes both apoptosis and necrosis, depending on dose and time. ATX activates cell death, stimulating the release of cytochrome C from mitochondria and the consequent activation of caspases-3. Moreover GM95 cells treated with ATX showed important lipidomic alterations, among them we detected a general decrease in several phospholipid species and important changes in lipids involved in programmed cell death e.g. ceramide. The data suggest two different mechanisms of cell death caused by ATX, one leading to (mainly saturated) glycerophospholipid hydrolysis related to an increase in diacylglycerols and associated to membrane damage and necrosis, and a second mechanism involving chiefly sphingomyelin hydrolysis and generation of proapoptotic lipidic mediators such as ceramide, N-acylethanolamine and saturated non-esterified fatty acids.
authors.affiliation: Unidad de Biofísica (CSIC, UPV/EHU), and Departamento de Bioquímica, Universidad del País Vasco, Aptdo. 644, 48080 Bilbao, Spain.
authors.forename: Marco M
authors.initials: MM
authors.lastname: Manni
authors.affiliation: Unidad de Biofísica (CSIC, UPV/EHU), and Departamento de Bioquímica, Universidad del País Vasco, Aptdo. 644, 48080 Bilbao, Spain.
authors.forename: Juan G
authors.initials: JG
authors.lastname: Valero
authors.affiliation: OWL, Parque Tecnológico de Bizkaia, Bizkaia, Spain.
authors.forename: Miriam
authors.initials: M
authors.lastname: Pérez-Cormenzana
authors.affiliation: OWL, Parque Tecnológico de Bizkaia, Bizkaia, Spain.
authors.forename: Ainara
authors.initials: A
authors.lastname: Cano
authors.affiliation: OWL, Parque Tecnológico de Bizkaia, Bizkaia, Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Alonso
authors.affiliation: Unidad de Biofísica (CSIC, UPV/EHU), and Departamento de Bioquímica, Universidad del País Vasco, Aptdo. 644, 48080 Bilbao, Spain. Electronic address: felix.goni@ehu.es.
authors.forename: Félix M
authors.initials: FM
authors.lastname: Goñi
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000737:chemistry / Q000633:toxicity*; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000633:toxicity*; D016923:Cell Death / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000737:chemistry / Q000633:toxicity*
chemical_list: D001427:Bacterial Toxins; D002135:Calcium-Binding Proteins; D008055:Lipids; D010738:Type C Phospholipases; C040649:alpha toxin, Clostridium perfringens
keywords: Alpha-toxin; Apoptosis; Ceramide; Diacylglycerol; GM95; Necrosis