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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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Book cover Polyamines

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1694))

Abstract

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.

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Acknowledgment

This work was supported by the Japan Society for the Promotion of Science (grant No. 26440144).

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Authors and Affiliations

  1. Graduate School of Biological Sciences, Nara Institute of Science and Technology, Takayama, 8916-5, Ikoma, Japan

    Tsubasa Shoji

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  1. Tsubasa Shoji
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Correspondence to Tsubasa Shoji .

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Editors and Affiliations

  1. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Rubén Alcázar

  2. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Antonio F. Tiburcio

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Shoji, T. (2018). 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. In: Alcázar, R., Tiburcio, A. (eds) Polyamines. Methods in Molecular Biology, vol 1694. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7398-9_20

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  • DOI: https://doi.org/10.1007/978-1-4939-7398-9_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7397-2

  • Online ISBN: 978-1-4939-7398-9

  • eBook Packages: Springer Protocols

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