Planta

, Volume 221, Issue 5, pp 690–704

Proteome analysis of the medicinal plant Catharanthus roseus

  • Denise I. Jacobs
  • Marco Gaspari
  • Jan van der Greef
  • Robert van der Heijden
  • Robert Verpoorte
Original Article

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.

Keywords

Alkaloid biosynthesis Catharanthus MALDI-MS/MS Proteomics Strictosidine synthase 

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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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Denise I. Jacobs
    • 1
  • Marco Gaspari
    • 2
  • Jan van der Greef
    • 2
  • Robert van der Heijden
    • 2
  • Robert Verpoorte
    • 1
  1. 1.Division of Pharmacognosy, Leiden/Amsterdam Center for Drug ResearchGorlaeus LaboratoriesLeidenThe Netherlands
  2. 2.Division of Analytical Biosciences, Leiden/Amsterdam Center for Drug ResearchGorlaeus LaboratoriesLeidenThe Netherlands

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