title: Mitochondrial Redox Opto-Lipidomics Reveals Mono-Oxygenated Cardiolipins as Pro-Apoptotic Death Signals.
abstract: While opto-genetics has proven to have tremendous value in revealing the functions of the macromolecular machinery in cells, it is not amenable to exploration of small molecules such as phospholipids (PLs). Here, we describe a redox opto-lipidomics approach based on a combination of high affinity light-sensitive ligands to specific PLs in mitochondria with LC-MS based redox lipidomics/bioinformatics analysis for the characterization of pro-apoptotic lipid signals. We identified the formation of mono-oxygenated derivatives of C18:2-containing cardiolipins (CLs) in mitochondria after the exposure of 10-nonylacridine orange bromide (NAO)-loaded cells to light. We ascertained that these signals emerge as an immediate opto-lipidomics response, but they decay long before the commencement of apoptotic cell death. We found that a protonophoric uncoupler caused depolarization of mitochondria and prevented the mitochondrial accumulation of NAO, inhibited the formation of C18:2-CL oxidation product,s and protected cells from death. Redox opto-lipidomics extends the power of opto-biologic protocols to studies of small PL molecules resilient to opto-genetic manipulations.
authors.forename: Gaowei
authors.initials: G
authors.lastname: Mao
authors.forename: Feng
authors.initials: F
authors.lastname: Qu
authors.forename: Claudette M
authors.initials: CM
authors.lastname: St Croix
authors.forename: Yulia Y
authors.initials: YY
authors.lastname: Tyurina
authors.affiliation: Division of Metabolic and Vascular Health, Medical School, University of Warwick , Coventry, United Kingdom.
authors.forename: Joan
authors.initials: J
authors.lastname: Planas-Iglesias
authors.forename: Jianfei
authors.initials: J
authors.lastname: Jiang
authors.forename: Zhentai
authors.initials: Z
authors.lastname: Huang
authors.forename: Andrew A
authors.initials: AA
authors.lastname: Amoscato
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Tyurin
authors.forename: Alexandr A
authors.initials: AA
authors.lastname: Kapralov
authors.forename: Amin
authors.initials: A
authors.lastname: Cheikhi
authors.forename: John
authors.initials: J
authors.lastname: Maguire
authors.affiliation: Division of Metabolic and Vascular Health, Medical School, University of Warwick , Coventry, United Kingdom.
authors.forename: Judith
authors.initials: J
authors.lastname: Klein-Seetharaman
authors.affiliation: Department of Critical Care Medicine, Safar Center for Resuscitation Research, University of Pittsburgh , Pittsburgh, Pennsylvania, United States.
authors.forename: Hülya
authors.initials: H
authors.lastname: Bayır
authors.forename: Valerian E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D000165:Acridine Orange / Q000031:analogs & derivatives / Q000378:metabolism; D017209:Apoptosis / Q000528:radiation effects; D002308:Cardiolipins / Q000737:chemistry / Q000378:metabolism*; D004396:Coloring Agents / Q000378:metabolism; D019295:Computational Biology; D006367:HeLa Cells; D006801:Humans; D008027:Light; D008928:Mitochondria / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002308:Cardiolipins; D004396:Coloring Agents; C056258:N(10)-nonylacridine orange; D000165:Acridine Orange; D010100:Oxygen