Takeshige K, Sumimoto H, Minakami S
Department of Biochemistry, Kyushu University School of Medicine, Fukuoka, Japan.
Biomed Biochim Acta. 1989;48(2-3):S25-30.
Polymorphonuclear leukocytes (PMNL) release superoxide anions formed by a membrane-bound NADPH oxidase induced by stimulations. Properties of the inducers and their antagonists indicate that Ca2+, GTP-binding protein (G-protein), phospholipase C and Ca2+, phospholipid-dependent protein kinase (C-kinase) are mainly associated with the stimulation of receptors. Low concentrations of ATP induce the oxidase accompanied by the increase in the intracellular Ca2+ due to the flux from the medium and the storage site. ATP-gamma-S, UTP and ITP are effective but mononucleotides, dinucleotides, GTP and CTP are not. Leukotriene B4 (LTB4) which acts as a chemotactic agent and the inducer of the NADPH oxidase is catabolized. It is hydroxylated by a specific cytochrome P450 and then oxidized to a carboxy derivative by a cytosolic alcohol dehydrogenase and a microsomal aldehyde dehydrogenase in PMNL. Active NADPH oxidase was obtained by incubating membrane and cytosolic components of resting PMNL in the presence of sodium dodecyl sulfate (SDS). Two cytosolic components were obtained by an affinity chromatography on 2',5'-ADP Sepharose. One component is active in the presence of GTP or GTP-gamma-S and the other component in the presence of another cytosolic fraction.
多形核白细胞（PMNL）释放由刺激诱导的膜结合NADPH氧化酶形成的超氧阴离子。诱导剂及其拮抗剂的特性表明，Ca2+、鸟苷三磷酸结合蛋白（G蛋白）、磷脂酶C和Ca2+、磷脂依赖性蛋白激酶（C激酶）主要与受体的刺激有关。低浓度的ATP诱导氧化酶，同时由于来自培养基和储存部位的通量导致细胞内Ca2+增加。ATP-γ-S、UTP和ITP有效，但单核苷酸、二核苷酸、GTP和CTP无效。作为趋化剂和NADPH氧化酶诱导剂的白三烯B4（LTB4）被分解代谢。它在PMNL中被一种特定的细胞色素P450羟基化，然后被细胞质乙醇脱氢酶和微粒体醛脱氢酶氧化为羧基衍生物。通过在十二烷基硫酸钠（SDS）存在下孵育静止PMNL的膜和细胞质成分获得活性NADPH氧化酶。通过在2',5'-ADP琼脂糖上进行亲和层析获得两种细胞质成分。一种成分在GTP或GTP-γ-S存在下有活性，另一种成分在另一种细胞质组分存在下有活性。
