Analysis of localization and function of the COOH-terminal corresponding site of cytochrome b558 in fish neutrophils.
Using an antibody against the synthetic peptide corresponding to the COOH-terminal region of human cytochrome b558 large subunit, a broad band was specifically detected in neutrophil lysates from 6 marine fish and 2 freshwater fish by western blotting. Immunofluorescence assay showed that the antibody recognized the epitopes in eel and tilapia neutrophils permeabilized with detergent. These results suggest that the cytochrome b large subunit universally exists in fish neutrophils and that the epitopes are exposed to the cytoplasmic side of fish neutrophils as well as human neutrophils. Furthermore, a synthetic peptide corresponding to the COOH-terminus of the large subunit apparently blocked superoxide production in a specific and dose-dependent fashion in eel and tilapia neutrophils, indicating that the region equivalent to the COOH-terminus of cytochrome b large subunit is responsible for superoxide generation in fish neutrophils.
Shiibashi
T
T
United Graduate School of Agricultural Sciences, Kagoshima University, Japan.
Iida
T
T
Itou
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003573:Cytochrome b Group / Q000378:metabolism / Q000502:physiology*; D005399:Fishes; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D015151:Immunoblotting; D008969:Molecular Sequence Data; D019255:NADPH Oxidases; D009504:Neutrophils / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism
D003573:Cytochrome b Group; D010455:Peptides; D013481:Superoxides; C036888:cytochrome b558; D019255:NADPH Oxidases