title: Biochemical characterization of reactive nitrogen species by eosinophil peroxidase in tyrosine nitration.
abstract: It is well known that eosinophils are involved in tyrosine nitration. In this study, we evaluated tyrosine nitration by rat eosinophils isolated from peritoneal fluid and constituent eosinophils in the stomach. Rat peritoneal eosinophils activated with 1 microM phorbol myristate acetate (PMA) and 50 microM NO2- showed immunostaining for nitrotyrosine only in smaller cells, despite the fact that eosinophils are capable of producing superoxide (O2*-) Free tyrosine nitrating capacity after incubation with PMA and NO2- was 4-fold higher in eosinophils than in neutrophils. Catalase and alpha- and gamma-tocopherol inhibited free tyrosine nitration by reactive nitrogen species from eosinophils but not that by peroxynitrite. Superoxide dismutase augmented free tyrosine nitration by activated eosinophils and peroxynitrite. The concentration of nitric oxide released from eosinophils was relatively low (0.32 microM/10(6) cells/h) and did not contribute to the formation of nitrotyrosine. On the other hand, most constituent eosinophils constituent in the rat stomach stimulated by PMA and NO2- showed tyrosine nitration capacity. These results suggest that intact cells other than apoptotic-like eosinophils eluted in the intraperitoneal cavity could not generate reactive species responsible for nitration by a peroxidase-dependent mechanism. In contrast, normal eosinophils in the stomach were capable of nitration, suggesting that the characteristics of eosinophils in gastric mucosa are different from those eluted in the peritoneal cavity.
authors.affiliation: Department of Viral Infection and International Health, Kanazawa University Graduate School of Medical Science, Kanazawa 920-0934, Japan.
authors.forename: Kei
authors.initials: K
authors.lastname: Takemoto
authors.forename: Keiki
authors.initials: K
authors.lastname: Ogino
authors.forename: Da-Hong
authors.initials: DH
authors.lastname: Wang
authors.forename: Tomoko
authors.initials: T
authors.lastname: Takigawa
authors.forename: Carmen M
authors.initials: CM
authors.lastname: Kurosawa
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Kamyabashi
authors.forename: Yuri
authors.initials: Y
authors.lastname: Hibino
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Hitomi
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ichimura
mesh_terms: D000818:Animals; D015153:Blotting, Western; D047209:Eosinophil Peroxidase / Q000378:metabolism*; D004804:Eosinophils / Q000187:drug effects / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology / Q000378:metabolism; D006651:Histocytochemistry; D007150:Immunohistochemistry; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D009569:Nitric Oxide / Q000494:pharmacology; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010537:Peritoneum / Q000166:cytology / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D026361:Reactive Nitrogen Species / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000096:biosynthesis
chemical_list: D012333:RNA, Messenger; D026361:Reactive Nitrogen Species; D013481:Superoxides; D009569:Nitric Oxide; C002744:3-nitrotyrosine; D014443:Tyrosine; D047209:Eosinophil Peroxidase; D052247:Nitric Oxide Synthase Type II; D013755:Tetradecanoylphorbol Acetate