Tetrameric structure and cellular location of catechol 2,3-dioxygenase
Tetrameric structure and cellular location of catechol 2,3-dioxygenase
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儿茶酚2,3-双加氧酶的四聚体结构和细胞定位
DOI：
10.1007/bf00262205
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发表时间：
2004
影响因子：
2.8
通讯作者：
M. Rohde
中科院分区：
文献类型：
--
作者：
J. Winkler;L. Eltis;D. Dwyer;M. Rohde
用电子显微镜研究了恶臭假单胞菌TOL质粒（pWWO）中邻苯二酚2，3-双加氧酶的结构。纯化的儿茶酚2，3-双加氧酶的负染样品显示，该酶由四个亚基排列在一个四面体构象。抗邻苯二酚2，3-双加氧酶的单克隆抗体显示出高度特异性反应，并用于大肠杆菌（pAW 31）和P. putida（pWWO），使用作为包埋后免疫电子显微术程序进行的蛋白A-金技术。我们的原位标记研究显示，在这两种细胞类型的儿茶酚2，3-双加氧酶的细胞质位置。
Catechol 2,3-dioxygenase from themeta-cleavage pathway encoded on the TOL plasmid ofPseudomonas putida(pWWO) was investigated by electron microscopy. Negatively stained samples of the purified catechol 2,3-dioxygenase revealed that the enzyme consists of four subunits arranged in a tetrahedral conformation. Monoclonal antibodies raised against catechol 2,3-dioxygenase showed highly specific reactions and were used to localize the enzyme inEscherichia coli(pAW31) andP. putida(pWWO), using the protein A-gold technique carried out as a post-embedding immunoelectron microscopy procedure. Our in situ labeling studies revealed a cytoplasmic location of the catechol 2,3-dioxygenase in both cell types.