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Detection of Phytochrome Phosphorylation in Plants

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Phytochromes

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2026))

Abstract

Posttranslational modification (PTM) of proteins occurs during or after translation and in most cases means covalent binding of a functional group to certain amino acid side chains. Among PTMs, phosphorylation is extensively studied for decades. During phosphorylation, a phosphate group is added to the target residue that is dominantly serine, threonine, and tyrosine in eukaryotes. The phosphate group attachment is catalyzed by kinases, whereas the removal of phosphate (dephosphorylation) is performed by phosphatases. Phosphorylation of phytochrome photoreceptors alters light signaling in multiple ways, thus the examination of this PTM is an expanding aspect of light signaling research. Although this chapter presents methods for detecting phosphorylated phytochrome B molecules, it can be applied on other phytochrome species. The first presented protocol of this chapter shows how the phosphorylation state of phytochrome photoreceptors can be monitored in a modified polyacrylamide gel electrophoresis system. The second protocol describes in detail how phosphorylated amino acids of a target molecule can be identified using mass spectrometry analysis.

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Acknowledgments

We thank Katalin F. Medzihradszky for critical reading of the manuscript. This work was supported by grants from the Economic Development and Innovation Operative Program (GINOP-2.3.2-15-2016-00015 and GINOP-2.3.2-15-2016-00032).

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Authors and Affiliations

  1. Laboratory of Proteomics Research, Biological Research Centre, Szeged, Hungary

    Eva Klement

  2. Agricultural Biotechnology Institute, National Agricultural Research and Innovation Centre, Gödöllő, Hungary

    Péter Gyula

  3. Plant Biology Institute, Biological Research Centre, Szeged, Hungary

    András Viczián

Authors
  1. Eva Klement
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  2. Péter Gyula
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  3. András Viczián
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Corresponding author

Correspondence to András Viczián .

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Editors and Affiliations

  1. Faculty of Biology, Institute of Biology II and Signalling Research Centres BIOSS and CIBSS, University of Freiburg, Freiburg, Germany

    Andreas Hiltbrunner

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Klement, E., Gyula, P., Viczián, A. (2019). Detection of Phytochrome Phosphorylation in Plants. In: Hiltbrunner, A. (eds) Phytochromes. Methods in Molecular Biology, vol 2026. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9612-4_4

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  • DOI: https://doi.org/10.1007/978-1-4939-9612-4_4

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9611-7

  • Online ISBN: 978-1-4939-9612-4

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