Plant Physiol.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via ISI Web of Science (18)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van Tegelen, L. J.P.
Right arrow Articles by Wullems, G. J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van Tegelen, L. J.P.
Right arrow Articles by Wullems, G. J.
Agricola
Right arrow Articles by van Tegelen, L. J.P.
Right arrow Articles by Wullems, G. J.

Purification and cDNA Cloning of Isochorismate Synthase from Elicited Cell Cultures of Catharanthus roseus

Léon J.P. van Tegelen, Paolo R.H. Moreno1, Anton F. Croes*, Robert Verpoorte, and George J. Wullems

Department of Experimental Botany, University of Nijmegen, Toernooiveld 1, 6525 ED Nijmegen, The Netherlands (L.J.P.v.T., A.F.C., G.J.W.); and Leiden/Amsterdam Center for Drug Research, Division of Pharmacognosy, Gorlaeus Laboratories, Leiden University, P.O. Box 9502, 2300 RA Leiden, The Netherlands (P.R.H.M., R.V.)

Isochorismate is an important metabolite formed at the end of the shikimate pathway, which is involved in the synthesis of both primary and secondary metabolites. It is synthesized from chorismate in a reaction catalyzed by the enzyme isochorismate synthase (ICS; EC 5.4.99.6). We have purified ICS to homogeneity from elicited Catharanthus roseus cell cultures. Two isoforms with an apparent molecular mass of 64 kD were purified and characterized. The Km values for chorismate were 558 and 319 µM for isoforms I and II, respectively. The isoforms were not inhibited by aromatic amino acids and required Mg2+ for enzyme activity. Polymerase chain reaction on a cDNA library from elicited C. roseus cells with a degenerated primer based on the sequence of an internal peptide from isoform II resulted in an amplification product that was used to screen the cDNA library. This led to the first isolation, to our knowledge, of a plant ICS cDNA. The cDNA encodes a protein of 64 kD with an N-terminal chloroplast-targeting signal. The deduced amino acid sequence shares homology with bacterial ICS and also with anthranilate synthases from plants. Southern analysis indicates the existence of only one ICS gene in C. roseus.

1   Present address: Department of Quinica Fundamental, Instituto de Quinica, Universida de São Paolo, São Paolo, Brazil.
*   Corresponding author; e-mail croes{at}sci.kun.nl; fax 31-24-3553450.

Plant Physiol. (1999) 119: 705-712
Copyright Clearance Center:   0032-0889/99/119//08
© 1999 American Society of Plant Physiologists




This article has been cited by other articles:


Home page
 Plant Physiol.Home page
C. Garcion, A. Lohmann, E. Lamodiere, J. Catinot, A. Buchala, P. Doermann, and J.-P. Metraux
Characterization and Biological Function of the ISOCHORISMATE SYNTHASE2 Gene of Arabidopsis
Plant Physiology, July 1, 2008; 147(3): 1279 - 1287.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. A. Strawn, S. K. Marr, K. Inoue, N. Inada, C. Zubieta, and M. C. Wildermuth
Arabidopsis Isochorismate Synthase Functional in Pathogen-induced Salicylate Biosynthesis Exhibits Properties Consistent with a Role in Diverse Stress Responses
J. Biol. Chem., February 23, 2007; 282(8): 5919 - 5933.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. Gaille, P. Kast, and D. Haas
Salicylate Biosynthesis in Pseudomonas aeruginosa. PURIFICATION AND CHARACTERIZATION OF PchB, A NOVEL BIFUNCTIONAL ENZYME DISPLAYING ISOCHORISMATE PYRUVATE-LYASE AND CHORISMATE MUTASE ACTIVITIES
J. Biol. Chem., June 7, 2002; 277(24): 21768 - 21775.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
ASPB Publications PLANT PHYSIOLOGY THE PLANT CELL
Copyright © 1999 by the American Society of Plant Biologists