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Suspension-Cultured Plant Cells as a Tool to Analyze the Extracellular Proteome

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Plant Proteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1072))

Abstract

Suspension-cultured cells (SCC) are generally considered the most suitable cell systems to carry out scientific studies, including the extracellular proteome (secretome). SCC are initiated by transferring friable callus fragments into flasks containing liquid culture medium for cell biomass growth, and they are maintained in an orbital shaker to supply the sufficient oxygen that allows cell growth. SCC increase rapidly during the exponential phase and after 10–20 days (depending on the cell culture nature), the growth rate starts to decrease due to limitation of nutrients, and to maintain for decades these kinds of cell cultures is needed to transfer a portion of these SCC into a fresh culture medium. Despite the central role played by extracellular proteins in most processes that control growth and development, the secretome has been less well characterized than other subcellular compartments, meaning that our understanding of the cell wall physiology is still very limited. Useful proteomic tools have emerged in recent years to unravel metabolic network that occurs in cell walls. With the recent progress made in mass spectrometry technology, it has become feasible to identify proteins from a given organ, tissue, cells, or even a subcellular compartment. Compared with other methods used to isolate cell wall proteins, the spent medium of SCC provides a convenient, continuous, and reliable and unique source of extracellular proteins. Therefore, this biological system could be used as a large-scale cell culture from which these proteins can be secreted, easily separated from cells without cell disruption, and so, without any cytosolic contamination, easily recovered from the extracellular medium. This nondestructive cell wall proteome approach discloses a set of proteins that are specifically expressed in the remodelling of the cell wall architecture and stress defense.

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Acknowledgements

This work has been supported by research grants from MICINN-FEDER (BIO2008-2941, BFU2009-08151, and BIO2011-29856-C02-02). ABSJ and AL held a grant from the Ministerio de Ciencia e Innovación. MJME acknowledges a grant from Fundación Caja Murcia-University of Alicante. University of Alicante Proteomics Facility is a member of Proteored (ISCIII). JCV acknowledges support from Red Temática de Investigacion Cooperativa de Cáncer (RTICC).

Author information

Authors and Affiliations

  1. Plant Peroxidases Group, Department of Plant Physiology, Faculty of Biology, University of Murcia, Murcia, Spain

    Ana B. Sabater-Jara, Lorena Almagro, Sarai Belchí-Navarro, Sabry M. Youssef, Juan C. Vera-Urbina, Susana Sellés-Marchart & María A. Pedreño

  2. Plant Proteomics and Functional Genomics Group, Department of Agrochemistry and Biochemistry, Faculty of Science, University of Alicante, Alicante, Spain

    María J. Martínez-Esteso & Roque Bru-Martínez

  3. Translational Oncology Unit, Instituto de Investigaciones Biomédicas ‘Alberto Sols’, Spanish National Research Council (CSIC-UAM), Madrid, Spain

    Juan Casado-Vela

Authors
  1. Ana B. Sabater-Jara
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  2. Lorena Almagro
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  3. Sarai Belchí-Navarro
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  4. María J. Martínez-Esteso
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  5. Sabry M. Youssef
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  6. Juan Casado-Vela
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  7. Juan C. Vera-Urbina
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  8. Susana Sellés-Marchart
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  9. Roque Bru-Martínez
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  10. María A. Pedreño
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Editor information

Editors and Affiliations

  1. Dept of Biochem & Molecular Bio, University of Cordoba, Cordoba, Spain

    Jesus V. Jorrin-Novo

  2. University of Tsukuba National Institute of Crop Science, Tsukuba, Japan

    Setsuko Komatsu

  3. Molecular Systems Biology, University of Vienna, Vienna, Austria

    Wolfram Weckwerth

  4. Dept. Molecular Systems Biology, University of Vienna, Vienna, Austria

    Stefanie Wienkoop

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Sabater-Jara, A.B. et al. (2014). Suspension-Cultured Plant Cells as a Tool to Analyze the Extracellular Proteome. In: Jorrin-Novo, J., Komatsu, S., Weckwerth, W., Wienkoop, S. (eds) Plant Proteomics. Methods in Molecular Biology, vol 1072. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-631-3_29

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  • DOI: https://doi.org/10.1007/978-1-62703-631-3_29

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-630-6

  • Online ISBN: 978-1-62703-631-3

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