Tsunawaki S, Mizunari H, Nagata M, Tatsuzawa O, Kuratsuji T
National Children's Medical Research Center, Tokyo, Japan.
Biochem Biophys Res Commun. 1994 Mar 30;199(3):1378-87. doi: 10.1006/bbrc.1994.1383.
Cytosolic components of human neutrophils, p47phox and p67phox, deficiencies of which lead to chronic granulomatous disease (CGD), potentiate respiratory burst oxidase translocating from cytosol to membrane upon cell stimulation. In this report we describe a novel cytosolic component, p40phox, which consistently behaves with p67phox through immunoprecipitation and column works, and is missing in patients with CGD who lack p67phox. Although actin has been reported to be involved in O2- generation, the p40phox profile did not correspond to that of actin. The tight association between p40phox and p67phox was not affected by treatment with a mixture of deoxycholate and Nonidet P-40, until subjected to SDS-PAGE. Addition of recombinant p67phox to cytosol did not produce any additional p40phox in the immunoprecipitate, unlike the additive increment in the band of p67phox. These results suggest that p40phox forms a complex with p67phox in a molar ratio of 1:1, without any free p40phox in the cytosol.
人类中性粒细胞的胞质成分p47phox和p67phox，其缺陷会导致慢性肉芽肿病（CGD），在细胞受到刺激时可增强呼吸爆发氧化酶从胞质溶胶向细胞膜的转运。在本报告中，我们描述了一种新的胞质成分p40phox，它通过免疫沉淀和柱层析操作与p67phox表现一致，且在缺乏p67phox的CGD患者中缺失。尽管有报道称肌动蛋白参与了超氧阴离子（O2-）的产生，但p40phox的分布与肌动蛋白的不同。在进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）之前，用脱氧胆酸盐和诺乃洗涤剂P-40的混合物处理并不会影响p40phox与p67phox之间的紧密结合。与p67phox条带的累加增加不同，向胞质溶胶中添加重组p67phox并不会在免疫沉淀物中产生任何额外的p40phox。这些结果表明，p40phox与p67phox以1:1的摩尔比形成复合物，胞质溶胶中不存在游离的p40phox。
