Abstract
The ability of a plant cytochrome P450 to bind and metabolise plant endogenous molecules and xenobiotics was investigated. The work was performed on the yeast-expressed CYP73A1, a cinnamate 4-hydroxylase isolated fromHelianthus tuberosus. CYP73 controls the general phenylpropanoid pathway and is likely to be one of the most abundant sources of P450 in the biosphere. The enzyme shows a high selectivity toward plant secondary metabolites. Nevertheless, it oxygenates several small and planar xenobiotics with low efficiency, including an herbicide (chlorotoluron). One xenobiotic molecule, 2naphthoic acid, is hydroxylated with an efficiency comparable to that of the physiological substrate. This reaction was used to devise a fluorimetric test for the rapid measurement of enzyme activity. A series of herbicidal molecules (hydroxybenzonitriles) are shown to bind the active site without being metabolised. These molecules behave as strong competitive inhibitors of CYP73 with a Ki in the same micromolar range as the Km for the physiological substrate. It is proposed that their inhibition of the phenylpropanoid pathway reinforces their other phytotoxic effects at the level of the chloroplasts. All our results indicate a strong reciprocal interaction between plant P450s and xenobiotics.
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D. Nelson;T. Kamataki;DJ. Waxman;F.P. Guengerich;R.W. Estabrook;R. Feyereisen;F.J. Gonzalez;M.J. Coon;I.C. Gunsalus;O. Gotoh;K. Okuda;D.S. Nebert: DNA Cell Biol. (1993)12, 1–51
R.L Lindberg;M. Negishi: Nature (1989)339, 632–634
F. Durst;I. Benveniste, in:Schenkman, J.B.;Greim, H. (eds) Handbook of experimental pharmacology (1993) Vol105, Springer Verlag, Heidelberg, pp 293–310
G.H. Teutsch;M.P. Hasenfratz;A. Lesot;C. Stoltz;J.M. Garnier;J.M. Jeltsch;F. Durst;D. Werck-Reichhart: Proc. Natl. Acad. Sci. USA (1993)90, 4102–4106
P. Urban;D. Werck-Reichhart;H.G. Teutsch;F. Durst;C. Mignotte;M. Kazmaier;D. Pompon: European J. Biochem. (1994)222, 843–850
G. Truan;C. Cullin;P. Reisdorf;P. Urban;D. Pompon: Gene (1993)125, 49–55
D. Werck-Reichhart;B. Gabriac;H. Teutsch;F. Durst: Biochem. J. (1990)270, 729–735
D. Reichhart;J.P. Salaün;I. Benveniste;F. Durst: Plant Physiol. (1980)66, 600–604
C. Grand: FEBS Letters (1984)169, 7–11
M.L. Hagmann;W. Heller;H. Grisebach: Eur. J. Biochem. (1983)134, 547–554
G. Kochs;H. Grisebach: Z., Naturforsch. (1987)42c, 343–348
G. Kochs;H. Grisebach: Eur. J. Biochem. (1986)155, 311–318
M.L. Hagmann;W. Heller;H. Grisebach: Eur. J. Biochem. (1984)142, 127–131
D. Hamerski;U. Matern: Eur.J. Biochem. (1988)171, 360–375
H. Wendorff;U. Matern: Eur J. Biochem. (1986)161, 391–398
D. Hamerski;U. Matern: FEBS Letters (1988)239, 263–265
H.P. Vetter;U. Mangold;G. Schröder;F.J. Marner;D. Werck-Reichhart;J. Schröder: Plant Physiol. (1992)100, 998–1007
J.P. Salaün;D. Weissbart;F. Durst;P. Pflieger;C. Mioskowski: FEBS Lett. (1989)246, 120–126
F. Pinot;J.P. Salaün;H. Bosch;A. Lesot;C. Mioskowski;F. Durst: Biochem. Biophys. Res. Commun. (1992)184, 183–193
A. Rahier;M. Taton: Biochem. Biophys. Res. Commun. (1986)140, 1064–1072
D.L. Hallahan;A.P. Heasman;M.C. Grossel;R. Quigley;P. Hedden;J.R. Bowyers: Plant Physiol. (1988)88, 1425–1429
A.B. Rifkind;T. J. Petschke: Pharmacol. Exp. Ther. (1981)217, 572–578
T. Nash: Biochem. J. (1953)55, 416–421
A. Zimmerlin;F. Durst: Plant Physiol. (1992)100, 874–881
J.J. McFadden;J.W. Gronwald;C.V. Eberlein: Biochem. Biophys. Res. Commun. (1990)168, 206–213
D. Reichhart;A. Simon;F. Durst;J.M. Mathews;P.R. Ortiz de Montellano: Archives Biochem. Biophys. (1982)216, 522–529
D. Werck-Reichhart;Y. Batard;G. Kochs;A. Lesot;F. Durst: Plant Physiol. (1993)102, 1291–1298
B. Gabriac;D. Werck-Reichhart;G.H. Teutsch;F. Durst: Arch. Biochem. Biophys. (1991)288, 302–309
I. Benveniste: Thèse de Doctorat d’Etat (1978) Université Louis Pasteur, Strasbourg
W. Oettmeier;C. Kude;H.J. Soll: Pestic. Biochem. Physiol. (1987)27, 50–60
D. Werck-Reichhart: Thèse de Doctorat d’Etat (1985) Université Louis Pasteur, Strasbourg
D.E. Schafer;D.O. Chilcote: Weed Sci. (1970)18, 729–732
A.E. Stapleton: Plant Cell (1992)4, 1353–1358
D. Reichhart;J.P. Salaün;I. Benveniste;F. Durst: Archives. Biochem. Biophys. (1979)196, 301–303
Y. Batard;A. Zimmerlin;M. Le Ret;F. Durst;D. Werck-Reichhart: Plant Cell Environm. (1995)18, 523–533
Y. Batard; M.A. Pierrel; A. Lesot; F. Durst; D. WerckReichhart: 2nd International Symposium on Cytochrome P450 of Microorganisms and Plants (1993) Tokyo
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Schalk, M., Pierrel, MA., Zimmerlin, A. et al. Xenobiotics: Substrates and inhibitors of the plant cytochrome P450. Environ. Sci. & Pollut. Res. 4, 229–234 (1997). https://doi.org/10.1007/BF02986353
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DOI: https://doi.org/10.1007/BF02986353