Abstract
5-Enolpyruvylshikimate 3-phosphate synthase (3-phosphoshikimate 1-carboxyvinyltransferase; EC 2.5.1.19) from shoot tissue of pea seedlings was purified to apparent homogeneity by sequential ammonium-sulphate precipitation, ion-exchange and hydrophobic-interaction chromatography and substrate elution from cellulose phosphate. Gel electrophoresis and gel-permeation chromatography showed that the purified enzyme was monomeric with molecular weight 50,000. The herbicide glyphosate was a potent inhibitor of the forward enzyme-catalyzed reaction.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- DEAE:
-
diethylaminoethyl
- EPSP:
-
5-enolpyruvylshikimate 3-phosphate
References
Ahmed, S.I., Giles, N.H. (1969) Organization of enzymes in the common aromatic synthetic pathway: evidence for aggregation in fungi. J. Bacteriol. 99, 231–237
Amrhein, N., Schab, J., Steinrücken, H.C. (1980) The mode of action of the herbicide glyphosate. Naturwissenschaften 67, 356–357
Berlyn, M.B., Ahmed, S.I., Giles, N.H. (1970) Organization of polyaromatic biosynthetic enzymes in a variety of photosynthetic organisms. J. Bacteriol. 104, 768–774
Berlyn, M.B., Giles, N.H. (1969) Organization of enzymes in the polyaromatic synthetic pathway: separability in bacteria. J. Bacteriol. 99, 222–230
Bode, R., Birnbaum, D. (1981a) Aggregation und Trennbarkeit der Enzyme des Shikimat-Pathway bei Hefen. Z. Allg. Mikrobiol. 21, 417–422
Bode, R., Birnbaum, D. (1981b) Eigenschaften und Regulation der Enzyme des Shikimat-Pathway von Hansenula henricii. Biochem. Physiol. Pflanz. 176, 329–341
Boocock, M.R., Coggins, J.R. (1983) Kinetics of 5-enolpyruvylshikimate-3-phosphate synthase inhibition by glyphosate. FEBS Lett. 154, 127–133
Boudet, A.M., Lecussan, R. (1974) Généralité de l'association (5-déshydroquinate hydro-lyase, shikimate: NADP+ oxydo-réductase) chez les végétaux supérieurs. Planta 119, 71–79
Boudet, A.M., Lecussan, R., Boudet, A. (1975) Characterization and properties of 3-dehydroquinate hydrolyases in higher plants. Planta 124, 67–76
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Burgoyne, L., Case, M.E., Giles, N.H. (1969) Purification and properties of the aromatic (arom) synthetic enzyme aggregate of Neurospora crassa. Biochim. Biophys. Acta 191, 452–462
Davis, B.J. (1964) Disc electrophoresis. II. Methods and applications to human serum proteins. Ann. N. Y. Acad. Sci. 121, 404–427
Gibson, F. (1970) Preparation of chorismic acid. Methods Enzymol. 17, 362–364
Gilchrist, D.G., Kosuge, T. (1980) Aromatic amino acid biosynthesis and its regulation. In: The biochemistry of plants, vol. 5: Amino acids and derivatives, pp. 507–531, Miflin, B.J., ed. Academic Press, New York London
Hasan, N., Nester, E.W. (1978) Dehydroquinate synthase in Bacillus subtilis: an enzyme associated with chorismate synthase and flavine reductase. J. Biol. Chem. 253, 4999–5004
Haslam, E. (1974) The shikimate pathway. Butterworths, London
Huang, L., Montoya, A.L., Nester, E.W. (1975) Purification and characterization of shikimate kinase enzyme activity in Bacillus subtilis. J. Biol. Chem. 250, 7675–7661
Koshiba, T. (1979) Purification of two forms of the associated 3-dehydroquinate hydro-lyase and shikimate: NADP+ oxidoreductase in Phaseolus mungo seedlings. Biochim. Biophys. Acta 522, 10–18
Koshiba, T. (1979) Organization of enzymes in the shikimate pathway of Phaseolus mungo seedlings. Plant Cell Physiol. 20, 667–670
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London) 224, 680–685
Lewendon, A., Coggins, J.R. (1983) Purification of 5-enolpyruvylshikimate 3-phosphate synthase from Escherichia coli. Biochem. J. 213, 187–191
Lumsden, J., Coggins, J.R. (1977) The subunit structure of the arom multienzyme complex of Neurospora crassa. Biochem. J. 161, 599–607
Patel, V.B., Giles, N.H. (1979) Purification of the arom multienzyme aggregate from Euglena gracilis. Biochim. Biophys. Acta 567, 24–34
Polley, L.D. (1978) Purification and characterisation of 3-dehydroquinate hydrolase and shikimate oxidoreductase. Biochim. Biophys. Acta 526, 259–266
Roisch, U., Lingens, F. (1980) Einfluß von N-(Phosphonomethyl)-glycin auf das Wachstum und auf die Enzyme der Aromatenbiosynthese von Escherichia coli. Hoppe-Seyler's Z. Physiol. Chem. 361, 1049–1058
Rubin, J.L., Gaines, C.G., Jensen, R.A. (1982) Enzymological basis for herbicidal action of glyphosate. Plant Physiol. 70, 833–839
Smith, D.D.S., Coggins, J.R. (1983) Isolation of a bifunctional domain from the pentafunctional arom enzyme complex of Neurospora crassa. Biochem. J. 213, 405–415
Steinrücken, H.C., Amrhein, N. (1980) The herbicide glyphosate is a potent inhibitor of 5-enolpyruvylshikimic acid-3-phosphate synthase. Biochem. Biophys. Res. Commun. 94, 1207–1212
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Mousdale, D.M., Coggins, J.R. Purification and properties of 5-enolpyruvylshikimate 3-phosphate synthase from seedlings of Pisum sativum L.. Planta 160, 78–83 (1984). https://doi.org/10.1007/BF00392469
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00392469