Abstract
Hydroxylation at the C-16 position of the indole alkaloid tabersonine has been suggested as the first step toward vindoline biosynthesis in Catharanthus roseus. Tabersonine 16-hydroxylase (16-OH) activity was detected in total protein extracts from young leaves of C. roseus using a novel coupled assay system. Enzyme activity was dependent on NADPH and molecular oxygen and was inhibited by CO, clotrimazole, miconazole, and cytochrome c. 16-OH was localized to the endoplasmic reticulum by linear sucrose density gradient centrifugation. These data suggest that 16-OH is a cytochrome P-450-dependent monooxygenase. The activity of 16-OH reached a maximum in seedlings 9 d postimbibition and was induced by light. The leaf-specific distribution of 16-OH in the mature plant is consistent with the localization of other enzymes in the tabersonine to vindoline pathway. However, in contrast to enzymes that catalyze the last four steps of vindoline biosynthesis, enzymes responsible for the first two steps from tabersonine (16-OH and 16-O-methyltransfersase) were detected in C. roseus cell-suspension cultures. These data complement the complex model of vindoline biosynthesis that has evolved with respect to enzyme compartmentalization, metabolic transport, and control mechanisms.
Full Text
The Full Text of this article is available as a PDF (1.5 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Aerts R. J., De Luca V. Phytochrome is involved in the light-regulation of vindoline biosynthesis in catharanthus. Plant Physiol. 1992 Oct;100(2):1029–1032. doi: 10.1104/pp.100.2.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Black S. D. Membrane topology of the mammalian P450 cytochromes. FASEB J. 1992 Jan 6;6(2):680–685. doi: 10.1096/fasebj.6.2.1537456. [DOI] [PubMed] [Google Scholar]
- De Carolis E., Chan F., Balsevich J., De Luca V. Isolation and Characterization of a 2-Oxoglutarate Dependent Dioxygenase Involved in the Second-to-Last Step in Vindoline Biosynthesis. Plant Physiol. 1990 Nov;94(3):1323–1329. doi: 10.1104/pp.94.3.1323. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Carolis E., De Luca V. Purification, characterization, and kinetic analysis of a 2-oxoglutarate-dependent dioxygenase involved in vindoline biosynthesis from Catharanthus roseus. J Biol Chem. 1993 Mar 15;268(8):5504–5511. [PubMed] [Google Scholar]
- De Luca V., Cutler A. J. Subcellular Localization of Enzymes Involved in Indole Alkaloid Biosynthesis in Catharanthus roseus. Plant Physiol. 1987 Dec;85(4):1099–1102. doi: 10.1104/pp.85.4.1099. [DOI] [PMC free article] [PubMed] [Google Scholar]
- De Luca V., Fernandez J. A., Campbell D., Kurz W. G. Developmental Regulation of Enzymes of Indole Alkaloid Biosynthesis in Catharanthus roseus. Plant Physiol. 1988 Feb;86(2):447–450. doi: 10.1104/pp.86.2.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Donaldson R. P., Luster D. G. Multiple forms of plant cytochromes p-450. Plant Physiol. 1991 Jul;96(3):669–674. doi: 10.1104/pp.96.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ESTABROOK R. W., COOPER D. Y., ROSENTHAL O. THE LIGHT REVERSIBLE CARBON MONOXIDE INHIBITION OF THE STEROID C21-HYDROXYLASE SYSTEM OF THE ADRENAL CORTEX. Biochem Z. 1963;338:741–755. [PubMed] [Google Scholar]
- Karp F., Mihaliak C. A., Harris J. L., Croteau R. Monoterpene biosynthesis: specificity of the hydroxylations of (-)-limonene by enzyme preparations from peppermint (Mentha piperita), spearmint (Mentha spicata), and perilla (Perilla frutescens) leaves. Arch Biochem Biophys. 1990 Jan;276(1):219–226. doi: 10.1016/0003-9861(90)90029-x. [DOI] [PubMed] [Google Scholar]
- Madyastha K. M., Ridgway J. E., Dwyer J. G., Coscia C. J. Subcellular localization of a cytochrome P-450-dependent monogenase in vesicles of the higher plant Catharanthus roseus. J Cell Biol. 1977 Feb;72(2):302–313. doi: 10.1083/jcb.72.2.302. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Meehan T. D., Coscia C. J. Hydroxylation of geraniol and nerol by a monooxygenase from Vinca rosea. Biochem Biophys Res Commun. 1973 Aug 21;53(4):1043–1048. doi: 10.1016/0006-291x(73)90570-6. [DOI] [PubMed] [Google Scholar]
- Meijer A. H., Souer E., Verpoorte R., Hoge J. H. Isolation of cytochrome P-450 cDNA clones from the higher plant Catharanthus roseus by a PCR strategy. Plant Mol Biol. 1993 May;22(2):379–383. doi: 10.1007/BF00014944. [DOI] [PubMed] [Google Scholar]
- Nebert D. W., Gonzalez F. J. P450 genes: structure, evolution, and regulation. Annu Rev Biochem. 1987;56:945–993. doi: 10.1146/annurev.bi.56.070187.004501. [DOI] [PubMed] [Google Scholar]
- Oshino N., Imai Y., Sato R. Electron-transfer mechanism associated with fatty acid desaturation catalyzed by liver microsomes. Biochim Biophys Acta. 1966 Oct 17;128(1):13–27. doi: 10.1016/0926-6593(66)90137-8. [DOI] [PubMed] [Google Scholar]
- Taniguchi H., Imai Y., Sato R. Role of the electron transfer system in microsomal drug monooxygenase reaction catalyzed by cytochrome P-450. Arch Biochem Biophys. 1984 Aug 1;232(2):585–596. doi: 10.1016/0003-9861(84)90577-0. [DOI] [PubMed] [Google Scholar]
- Teutsch H. G., Hasenfratz M. P., Lesot A., Stoltz C., Garnier J. M., Jeltsch J. M., Durst F., Werck-Reichhart D. Isolation and sequence of a cDNA encoding the Jerusalem artichoke cinnamate 4-hydroxylase, a major plant cytochrome P450 involved in the general phenylpropanoid pathway. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4102–4106. doi: 10.1073/pnas.90.9.4102. [DOI] [PMC free article] [PubMed] [Google Scholar]
- White R. E., Coon M. J. Oxygen activation by cytochrome P-450. Annu Rev Biochem. 1980;49:315–356. doi: 10.1146/annurev.bi.49.070180.001531. [DOI] [PubMed] [Google Scholar]