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Visual Mapping of Cell Wall Biosynthesis

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The Plant Cell Wall

Part of the book series: Methods in Molecular Biology ((MIMB,volume 715))

Abstract

Biosynthesis of pectin and hemicelluloses occurs in the Golgi apparatus and is thought to involve spatial regulations and complex formation of biosynthetic enzymes and proteins. We have demonstrated that a combination of heterologous expression of recombinant proteins tagged with fluorescent proteins and live cell imaging with confocal laser scanning microscopy (CLSM) allows efficient visualization of biosynthetic enzymes and proteins in subcellular compartments. We have also successfully utilized bimolecular fluorescence complementation (BiFC) for in situ visualization of protein–protein interactions of pectin biosynthetic enzymes and for the determination of their membrane topology in the Golgi apparatus.

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Acknowledgements

This work was supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research, through contract DE-AC02-05CH11231 between Lawrence Berkeley National Laboratory and the U.S. Department of Energy; by the Danish Villum Kann Rasmussen Foundation through the VKR centre Pro-active Plants; and by the Danish Agency for Science, Technology and Innovation.

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Authors and Affiliations

  1. The Department of Plant Biology and Biotechnology, University of Copenhagen, Copenhagen, Denmark

    Yumiko Sakuragi & Morten H. H. Nørholm

  2. Joint Bioenergy Institution, Feedstocks Division, Lawrence Berkeley National Laboratory, Emeryville, CA, USA

    Henrik V. Scheller

Authors
  1. Yumiko Sakuragi
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  2. Morten H. H. Nørholm
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  3. Henrik V. Scheller
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Editor information

Editors and Affiliations

  1. School of Natural Sciences, Dept. of Botany & Plant Science, National University of Ireland, Galway, University Road, Galway, Ireland

    Zoë A. Popper

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Sakuragi, Y., Nørholm, M.H.H., Scheller, H.V. (2011). Visual Mapping of Cell Wall Biosynthesis. In: Popper, Z. (eds) The Plant Cell Wall. Methods in Molecular Biology, vol 715. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-008-9_11

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  • DOI: https://doi.org/10.1007/978-1-61779-008-9_11

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-007-2

  • Online ISBN: 978-1-61779-008-9

  • eBook Packages: Springer Protocols

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