title: Random versus selective membrane phospholipid oxidation in apoptosis: role of phosphatidylserine.
abstract: The formation of reactive oxygen species has been associated with apoptosis. To assess the role of lipid peroxidation in apoptosis, we used 2,2'-azobis(2,4-dimethylisovaleronitrile) (AMVN) to generate peroxyl radicals within cellular membranes of HL-60 cells. cis-Parinaric acid (cis-PnA) metabolically integrated into phospholipids of HL-60 cells was used as a probe to assess the extent of lipid peroxidation within specific phospholipid classes. Within 2 h, AMVN (500 microM) randomly oxidized more than 85% of cis-PnA contained in all major classes of phospholipids. AMVN-induced lipid peroxidation was followed by apoptosis as determined by nuclear condensation, DNA fragmentation, and annexin V binding to externalized phosphatidylserine (PS). Fluorescamine derivatization of external aminophospholipids revealed that PS, but not phosphatidylethanolamine, was externalized. The vitamin E analogue, 6-hydroxy-2,2,5,7,8-pentamethylchromane (PMC), inhibited overall oxidation of cis-PnA in phospholipids by more than 85%. Not all phospholipids, however, were equally protected. Phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and sphingomyelin were nearly completely protected by PMC, while oxidation of PS was unaffected in whole living cells. The insensitivity of PS to PMC was not an intrinsic property because PMC protected all lipids equally during AMVN oxidation of liposomes prepared from cis-PnA-labeled cells. The potential role for PS oxidation in apoptosis was further suggested by the faithful execution of apoptosis following coexposure of cells to AMVN and PMC.
authors.affiliation: Department of Environmental and Occupational Health, School of Public Health, University of Pittsburgh, Pennsylvania 15238, USA.
authors.forename: J P
authors.initials: JP
authors.lastname: Fabisiak
authors.forename: Y Y
authors.initials: YY
authors.lastname: Tyurina
authors.forename: V A
authors.initials: VA
authors.lastname: Tyurin
authors.forename: J S
authors.initials: JS
authors.lastname: Lazo
authors.forename: V E
authors.initials: VE
authors.lastname: Kagan
mesh_terms: D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001391:Azo Compounds / Q000494:pharmacology; D002839:Chromans / Q000494:pharmacology; D004351:Drug Resistance; D005231:Fatty Acids, Unsaturated / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects; D008563:Membrane Lipids / Q000378:metabolism*; D009570:Nitriles / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D010718:Phosphatidylserines / Q000378:metabolism / Q000502:physiology*; D010743:Phospholipids / Q000378:metabolism*
chemical_list: D000975:Antioxidants; D001391:Azo Compounds; D002839:Chromans; D005231:Fatty Acids, Unsaturated; D008563:Membrane Lipids; D009570:Nitriles; D010718:Phosphatidylserines; D010743:Phospholipids; C055792:2,2'-azobis(2,4-dimethylvaleronitrile); C029141:2,2,5,7,8-pentamethyl-1-hydroxychroman; C013641:parinaric acid