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Proteome analysis of the medicinal plant Catharanthus roseus

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Abstract

A proteomic approach is undertaken aiming at the identification of novel proteins involved in the alkaloid biosynthesis of Catharanthus roseus. The C. roseus cell suspension culture A11 accumulates the terpenoid indole alkaloids strictosidine, ajmalicine and vindolinine. Cells were grown for 21 days, and alkaloid accumulation was monitored during this period. After a rapid increase between day 3 and day 6, the alkaloid content reached a maximum on day 16. Systematic analysis of the proteome was performed by two-dimensional polyacrylamide gel electrophoresis. After day 3, the proteome started to change with an increasing number of protein spots. On day 13, the proteome changed back to roughly the same as at the start of the growth cycle. 88 protein spots were selected for identification by mass spectrometry (MALDI-MS/MS). Of these, 58 were identified, including two isoforms of strictosidine synthase (EC 4.3.3.2), which catalyzes the formation of strictosidine in the alkaloid biosynthesis; tryptophan synthase (EC 4.1.1.28), which is needed for the supply of the alkaloid precursor tryptamine; 12-oxophytodienoate reductase, which is indirectly involved in the alkaloid biosynthesis as it catalyzes the last step in the biosynthesis of the regulator jasmonic acid. Unique sequences were found, which may also relate to unidentified biosynthetic proteins.

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Abbreviations

IPG:

Immobilized pH gradient

2D-PAGE:

Two-dimensional polyacrylamide gel electrophoresis

MALDI-TOF:

Matrix assisted laser desorption ionization time of flight

MS:

Mass spectrometry

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Acknowledgements

The authors would like to thank the Van Leersumfonds (the Netherlands) to make the purchase of an IPGphor and MALDI-sample plates possible.

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Authors and Affiliations

  1. Division of Pharmacognosy, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, P.O. Box 9502, 2300, Leiden, The Netherlands

    Denise I. Jacobs & Robert Verpoorte

  2. Division of Analytical Biosciences, Leiden/Amsterdam Center for Drug Research, Gorlaeus Laboratories, P.O. Box 9502, 2300, Leiden, The Netherlands

    Marco Gaspari, Jan van der Greef & Robert van der Heijden

Authors
  1. Denise I. Jacobs
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  2. Marco Gaspari
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  3. Jan van der Greef
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  4. Robert van der Heijden
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  5. Robert Verpoorte
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Correspondence to Robert van der Heijden.

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Jacobs, D.I., Gaspari, M., van der Greef, J. et al. Proteome analysis of the medicinal plant Catharanthus roseus. Planta 221, 690–704 (2005). https://doi.org/10.1007/s00425-004-1474-4

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  • DOI: https://doi.org/10.1007/s00425-004-1474-4

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