Abstract
Plants produce a wide variety of secondary metabolites (SMs) of pharmacological interest that, due to their high structural complexity, are difficult to obtain from chemical synthesis. Knowledge of the enzymatic process involved in the biosynthesis of these SMs may contribute to the development of new strategies for obtaining them from plant tissues. Maytenus ilicifolia contains three main classes of bioactive compounds: sesquiterpene pyridine alkaloids, flavonoids and, primarily, quinonemethide triterpenes, which are well known for their high and broad antitumor activity. Thus, to determine the enzymatic composition in the secondary metabolism of M. ilicifolia, we carried out proteomic profiling of M. ilicifolia cell cultures with and without increased concentrations of maytenin and 22β-hydroxy-maytenin by elicitation with methyl jasmonate for 48 h. For this purpose, the analysis of soluble proteins was performed via LCMS/MS experiments using a shotgun strategy and compared to specific databases, resulting in 1319 identifications in the two types of cells. Many enzymes involved in secondary metabolism were detected, including cytochrome P450-dependent monooxygenases, which potentially catalyze the final steps in the quinonemethide triterpene biosynthesis. In conclusion, we report for the first time a proteomic study of a cell culture system that is an effective elicitor of quinonemethide triterpene production.
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Acknowledgements
We thank the São Paulo Research Foundation (FAPESP) for the CIBFar-2013/07600-3 grant. T. A. Paz thanks CAPES for the provision of a scholarship. V. A. F. F. M. dos Santos and M. C. Inácio thank FAPESP for their fellowships (2011/01544-9 and 2014/19362-2, respectively). V. A. F. F. M. dos Santos is also grateful to the National Council for Scientific and Technological Development (CNPq) for her fellowship. M. Furlan and M. S. Palma would also like to thank CNPq for their research fellowships.
Author contributions
TAP was responsible for the conception and design of all experiments, data analysis, drafting and editing of the manuscript. MCI contributed with plant cell experiments and revision of the manuscript. VAFFMS contributed with proteomic experiments. NBD and MSP contributed with proteomic experiments and revised the manuscript. AMSP supervised part of experiments and contributed to data interpretation and final revision of the manuscript. MF contributed to the experimental design, to data interpretation, drafting and final revision of the manuscript, and supervised all experiments.
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Communicated by Ali R. Alan.
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Paz, T.A., dos Santos, V.A.F.F.M., Inácio, M.C. et al. Proteome profiling reveals insights into secondary metabolism in Maytenus ilicifolia (Celastraceae) cell cultures producing quinonemethide triterpenes. Plant Cell Tiss Organ Cult 130, 405–416 (2017). https://doi.org/10.1007/s11240-017-1236-1
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DOI: https://doi.org/10.1007/s11240-017-1236-1