Fuchs A, Dagher M C, Vignais P V
Commissariat à l'Energie Atomique/Laboratoire de Biochimie (URA 1130/CNRS), Département de Biologie Moléculaire et Structurale, Centre d'Etudes Nucléaires de Grenoble, France.
J Biol Chem. 1995 Mar 17;270(11):5695-7. doi: 10.1074/jbc.270.11.5695.
The superoxide-generating NADPH oxidase complex in phagocytic cells is constituted of a heterodimeric flavocytochrome b and cytosolic factors, p67phox, p47phox and p40phox as well as a small G protein Rac (for review, see Refs. 1-3). A truncated form of the p40phox cDNA was isolated by a two hybrid screen of a B lymphocyte library using a full length clone of p47phox as target. This truncated form of p40phox consisting of the Src Homology 3 (SH3) domain to the 3' stop codon was also shown to interact with p67phox in the same system. A library of smaller fragments of the truncated p40 cDNA was constructed and screened against either p47phox or p67phox. Results show that the SH3 domain of p40phox is sufficient for interaction with p47phox, whereas the C terminus of p40phox but not its SH3 domain is involved in the interaction with p67phox.
吞噬细胞中产生超氧化物的NADPH氧化酶复合物由异二聚体黄素细胞色素b和细胞溶质因子、p67phox、p47phox和p40phox以及小G蛋白Rac组成（综述见参考文献1 - 3）。使用p47phox的全长克隆作为靶标，通过对B淋巴细胞文库的双杂交筛选，分离出p40phox cDNA的截短形式。在同一系统中，这种由Src同源结构域3（SH3）至3'端终止密码子组成的p40phox截短形式也显示出与p67phox相互作用。构建了截短的p40 cDNA较小片段的文库，并针对p47phox或p67phox进行筛选。结果表明，p40phox的SH3结构域足以与p47phox相互作用，而p40phox的C末端而非其SH3结构域参与与p67phox的相互作用。
