Ohshima H, Brouet I M, Bandaletova T, Adachi H, Oguchi S, Iida S, Kurashima Y, Morishita Y, Sugimura T, Esumi H
Biochemistry Division, National Cancer Center Research Institute, Tokyo, Japan.
Biochem Biophys Res Commun. 1992 Sep 30;187(3):1291-7. doi: 10.1016/0006-291x(92)90443-o.
A polyclonal antibody was raised in the rabbit against an inducible form of nitric oxide (NO) synthase (EC 1.14.23) purified from the liver of rats with acute liver necrosis induced by i.v. administration of Propionibacterium acnes and lipopolysaccharide. The antibody immunoprecipitated NO synthase activities in the soluble extract of the liver from treated rats. Western blot analysis showed that the cytosols of the liver, lung and spleen from the treated rats but not from non-treated rats, and that of murine macrophages cultured in the presence of lipopolysaccharide and interferon-gamma, contained immunoreactive protein with a molecular weight of 125 kDa. The antibody, however, does not cross-react with a 150 kDa constitutive form of NO synthase present in the brain of rats, indicating that the inducible and constitutive enzymes are immunologically distinguishable.
用从经静脉注射痤疮丙酸杆菌和脂多糖诱导急性肝坏死的大鼠肝脏中纯化的诱导型一氧化氮（NO）合酶（EC 1.14.23），在兔体内制备了多克隆抗体。该抗体免疫沉淀了处理过的大鼠肝脏可溶性提取物中的NO合酶活性。蛋白质免疫印迹分析表明，处理过的大鼠而非未处理的大鼠的肝脏、肺和脾脏的胞质溶胶，以及在脂多糖和γ干扰素存在下培养的小鼠巨噬细胞的胞质溶胶，含有分子量为125 kDa的免疫反应性蛋白。然而，该抗体与大鼠脑中存在的150 kDa组成型NO合酶不发生交叉反应，表明诱导型和组成型酶在免疫上是可区分的。
