title: Peroxynitrite induces apoptosis of HL-60 cells by activation of a caspase-3 family protease.
abstract: Apoptosis is an active process critical for the homeostasis of organisms. Enzymes of the caspase family are responsible for executing this process. We have previously shown that peroxynitrite (ONOO-), a biological product generated from the interaction of nitric oxide and superoxide, induces apoptosis of HL-60 cells. The aim of this study was to elucidate the mechanisms involved in the execution process of peroxynitrite-induced apoptosis. Proteolytic cleavage of poly(ADP-ribose) polymerase, an indication of caspase-3 family protease activation and an early biochemical event accompanying apoptosis, was observed in a time-dependent manner during peroxynitrite-induced apoptosis of HL-60 cells. Activation of caspase-3 during peroxynitrite-induced apoptosis was substantiated by monitoring proteolysis of the caspase-3 proenzyme and by measuring caspase-3 activity with a fluorogenic substrate. Furthermore, pretreatment of HL-60 cells with N-acetyl-Asp-Glu-Val-Asp-aldehyde, a specific inhibitor of caspase-3, but not N-acetyl-Tyr-Val-Ala-Asp-aldehyde, a specific inhibitor of caspase-1, decreased peroxynitrite-induced apoptosis. These results suggest that the activation of a caspase-3 family protease is essential for initiating the execution process of peroxynitrite-induced apoptosis of HL-60 cells.
authors.affiliation: Department of Cell Biology, School of Osteopathic Medicine, University of Medicine and Dentistry of New Jersey, Stratford 08084, USA.
authors.forename: K T
authors.initials: KT
authors.lastname: Lin
authors.forename: J Y
authors.initials: JY
authors.lastname: Xue
authors.forename: M C
authors.initials: MC
authors.lastname: Lin
authors.forename: E G
authors.initials: EG
authors.lastname: Spokas
authors.forename: F F
authors.initials: FF
authors.lastname: Sun
authors.forename: P Y
authors.initials: PY
authors.lastname: Wong
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004789:Enzyme Activation / Q000502:physiology; D018922:HL-60 Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D009566:Nitrates / Q000494:pharmacology*; D016877:Oxidants / Q000494:pharmacology*; D010447:Peptide Hydrolases / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism
chemical_list: D015853:Cysteine Proteinase Inhibitors; D009566:Nitrates; D016877:Oxidants; C039642:peroxynitric acid; D011065:Poly(ADP-ribose) Polymerases; D010447:Peptide Hydrolases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases