Immunological studies on the respiratory burst oxidase of pig blood neutrophils

FEBS Letters

Fukuhara, Yukiko¹  Kakinuma, Katsuko¹  Ise, Yayoi¹ 

[1]Tokyo Metropolitan Institute of Medical Science, 18-22, Honkomagome 3-chome, Bunkyo-ku, Tokyo 113, Japan

关键词: Neutrophil;    Respiratory burst;    NADPH oxidase;    Superoxide generation;    FAD enzyme;    Antibody;    IEF—PAGE;    isoelectric focusing-polyacrylamide gel electrophoresis;    SDS—PAGE;    SDS-polyacrylamide gel electrophoresis;    NBT;    nitroblue tetrazolium;   

DOI  :  10.1016/0014-5793(88)80816-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Recently, a flavin enzyme (pI 5.0), that is probably responsible for superoxide (O⁻ ₂)-generating oxidase activity, was separated by isoelectric focusing-polyacrylamide gel electrophoresis (IEF—PAGE) from neutrophil membranes in our laboratory [(1987) J. Biol. Chem. 262, 12316–12322]. In the present work, we performed immunological studies on this enzyme derived from pig blood neutrophils. The enzyme extract obtained on IEF—PAGE was injected into guinea pigs to raise antibodies. IgG antibody against the pI 5.0 protein inhibited maximally 54% of the O⁻ ₂-generating activity of the membrane-solubilized oxidase, whereas the normal serum IgG was not inhibitory at all. Our results further confirmed that the enzyme (pI 5.0) is one of the component(s) of the O⁻ ₂-generating system. The enzyme gave rise to a band corresponding to a major protein of 72±4 kDa on both non-denaturing and SDS—PAGE. Immunoblotting after SDS—PAGE demonstrated labelling of peptides of 70–72, 28–32 and 16–18 kDa.

【 授权许可】

Unknown