Immunocytochemical localization of membrane-bound ammonia monooxygenase in cells of ammonia oxidizing bacteria
Immunocytochemical localization of membrane-bound ammonia monooxygenase in cells of ammonia oxidizing bacteria
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DOI：
10.1007/s00203-005-0074-4
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发表时间：
2006-03-01
影响因子：
2.8
通讯作者：
Bock, E
中科院分区：
文献类型：
--
作者：
Fiencke, C;Bock, E
用电子显微镜免疫细胞化学方法研究了膜结合型氨单加氧酶(AMO)在所有氨氧化细菌属(亚硝化单胞菌、亚硝球菌属和亚硝螺旋菌属)中的胞内定位。用识别AmoA和AmoB两个亚基的多克隆抗体进行包埋后标记。超薄切片显示AmoB蛋白定位于细胞膜上的所有属。在亚硝化单胞菌和亚硝化球菌的细胞中，胞质内膜上有额外的但较少的AmoB标记。与检测AmoB蛋白相反，AmoA抗体未能检测到AmoA蛋白。基于定量免疫印迹，亚硝酸单胞菌中ICM的范围与细胞内AMOA的含量相关。当底物浓度较低时，细菌的ICM和AMOA含量最大。
The intracellular location of the membrane-bound ammonia monooxygenase (AMO) in all genera of ammonia oxidizing bacteria (Nitrosomonas, Nitrosococcus and Nitrosospira) was determined by electron microscopic immunocytochemistry. Polyclonal antibodies recognizing the two subunits, AmoA- and AmoB-proteins, were used for post-embedding labeling. Ultrathin sections revealed that the AmoB-protein was located in all genera on the cytoplasmic membrane. In cells of Nitrosomonas and Nitrosococus additional but less AmoB-labeling was found on the intracytoplasmic membrane (ICM). In contrast to the detection of AmoB-protein, the AmoA-antibodies failed to detect the AmoA-protein. Based on quantitative immunoblots the extent of ICM in Nitrosomonas eutropha was correlated with the amount of AmoA in the cells. The highest extent of ICM and amount of AmoA was found at low ammonium substrate concentrations.