Achacin induces cell death in HeLa cells through two different mechanisms.
Achacin, which belongs to the L-amino acid oxidase group, oxidizes free amino acids and produces hydrogen peroxide in cell culture systems. Morphological changes in cells incubated with achacin were similar to those of cells incubated with H(2)O(2). In both cases, the end result was cell death. To examine the mechanism of achacin-associated cytotoxicity, the H(2)O(2) scavenger catalase was added to culture media. Features typical of apoptosis, including morphological changes, DNA fragmentation, and PARP cleavage, were observed when cells were incubated with achacin in the presence of catalase. Moreover, apoptosis was inhibited by Z-VAD-fmk, a broad-spectrum caspase inhibitor. Herein, we present evidence that two pathways are involved in achacin-induced cell death. One is direct generation of H(2)O(2) through the L-amino acid oxidase activity of achacin. The other is the caspase-mediated apoptotic pathway that is induced by depletion of L-amino acids by achacin.
Kanzawa
Nobuyuki
N
Department of Chemistry, Faculty of Science and Technology, Sophia University, 102-8554 Tokyo, Japan. n-kanza@sophia.ac.jp
Shintani
Satoko
S
Ohta
Kazumasa
K
Kitajima
Seiji
S
Ehara
Tatsuya
T
Kobayashi
Hiroko
H
Kizaki
Harutoshi
H
Tsuchiya
Takahide
T
D000590:Amino Acid Chloromethyl Ketones / Q000494:pharmacology; D000594:Amino Acid Oxidoreductases / Q000378:metabolism; D000596:Amino Acids / Q000032:analysis / Q000737:chemistry / Q000172:deficiency / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D002875:Chromosomes / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D050830:L-Amino Acid Oxidase; D009479:Neuropeptides / Q000378:metabolism / Q000494:pharmacology*; D010084:Oxidation-Reduction
D000590:Amino Acid Chloromethyl Ketones; D000596:Amino Acids; D061945:Caspase Inhibitors; D015853:Cysteine Proteinase Inhibitors; D009479:Neuropeptides; C096713:benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone; C077200:achacin; D006861:Hydrogen Peroxide; D000594:Amino Acid Oxidoreductases; D050830:L-Amino Acid Oxidase; D020169:Caspases