Analysis of the Intracellular Localization of Transiently Expressed and Fluorescently Labeled Copper-Containing Amine Oxidases, Diamine Oxidase and N-Methylputrescine Oxidase in Tobacco, Using an Agrobacterium Infiltration Protocol
Analysis of the Intracellular Localization of Transiently Expressed and Fluorescently Labeled Copper-Containing Amine Oxidases, Diamine Oxidase and N-Methylputrescine Oxidase in Tobacco, Using an Agrobacterium Infiltration Protocol
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DOI：
10.1007/978-1-4939-7398-9_20
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Shoji, Tsubasa
中科院分区：
文献类型：
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作者：
Shoji, Tsubasa
酶的细胞内定位为理解复杂的代谢途径提供了关键信息。基于酶的定位数据，多个细胞器的参与和细胞室之间的代谢物的运动已被建议为一些途径。通过农杆菌渗透在本氏烟草（Nicotiana benthamiana）叶片中瞬时表达荧光标记蛋白是检查感兴趣蛋白质的细胞内定位的简单且通用的方法。在这里，这种方法被施加到证明过氧化物酶体定位的同源含铜胺氧化酶（CuAOs）从烟草具有不同的底物偏好：二胺氧化酶（DAO），介导多胺catalysts，和N-甲基腐胺氧化酶（MPO），这是参与尼古丁的生物合成。我们的研究结果表明，农杆菌渗透协议可以有效地用于研究细胞内定位的过氧化物酶体的氧化酶定位。
The intracellular localization of enzymes provides key information for understanding complex metabolic pathways. Based on enzyme localization data, the involvement of multiple organelles and the movement of metabolites between cellular compartments have been suggested for a number of pathways. Transient expression of fluorescently tagged proteins in the leaves of Nicotiana benthamiana through Agrobacterium infiltration is a simple and versatile way to examine the intracellular localization of proteins of interest. Here, this method was applied to demonstrate the peroxisomal localization of a pair of homologous copper-containing amine oxidases (CuAOs) from tobacco with distinct substrate preferences: diamine oxidase (DAO), which mediates polyamine catabolism, and N-methylputrescine oxidase (MPO), which is involved in nicotine biosynthesis. Our results demonstrate that the Agrobacterium infiltration protocol can be effectively used to study the intracellular localization of oxidases that localize to the peroxisome.