Journal Article

Insect cell expression of recombinant imidazoleglycerolphosphate dehydratase of arabidopsis and wheat and inhibition by triazole herbicides

Tada, S. (Ciba-Geigy Japan Ltd., Takarazuka, Japan.), Guyer, D, Nakayama, Y, Hatano, M, Ohta, D, Ward, E, Volrath, S

1995

Overview

Imidazoleglycerolphosphate dehydratase (IGPD; EC 4.2.1.19), which is involved in the histidine biosynthetic pathway of Arabidopsis thaliana and wheat (Triticum aestivum), has been expressed in insect cells using the baculovirus expression vector system. N-terminal amino acid sequencing indicated that recombinant IGPDs (rlGPDs) were produced as mature forms via nonspecific proteolytic cleavages in the putative transit peptide region. The wheat rlGPD contained one Mn atom per subunit, and the Mn was involved in the assembly of the subunits to form active IGPDs. Protein-blotting analysis, using antibodies raised against the wheat rlGPD, indicated that IGPD was located in the chloroplasts of wheat. The rlGPDs of Arabidopsis and wheat, which were 86% identical in their primary structures deduced from the cDNAs, exhibited similar properties in terms of the molecular mass, pH optimum, and the Km for the substrate, imidazoleglycerolphosphase. However, the nonselective herbicides 3-amino-1,2,4-triazole and a newly synthesized triazole [(1R*, 3R*)-[3-hydroxy-3-(2H- [1,2,4]triazole-3-yl)-cyclohexyl]-phosphonic acid], inhibited Arabidopsis and wheat IGPDs in a mixed-type and a competitive manner respectively

• Publisher: American Society of Plant Physiologists
• Subject: ADN / ADN RECOMBINADO / ADN RECOMBINE / Amino Acid Sequence / amino acid sequences / Amitrole / Amitrole - pharmacology / Animals / antagonists & inhibitors / Arabidopsis / Arabidopsis - enzymology / Arabidopsis - genetics / ARABIDOPSIS THALIANA / AZOLE / AZOLES / BACULOVIRIDAE / Baculoviridae - genetics / Base Sequence / Biochemistry and Enzymology / Biological and medical sciences / BIOSINTESIS / BIOSYNTHESE / biosynthesis / Cell Line / Chemical control / CHLOROPLASTE / Chloroplasts / CLOROPLASTO / Complementary DNA / COMPOSICION QUIMICA / COMPOSITION CHIMIQUE / DNA, Plant / DNA, Plant - genetics / ELECTROFORESIS / ELECTROPHORESE / Enzyme Inhibitors / Enzyme Inhibitors - pharmacology / Enzymes / enzymology / EXPRESION GENICA / EXPRESSION DES GENES / Fundamental and applied biological sciences. Psychology / Gene Expression / genetic vectors / genetics / Herbicides / Herbicides - pharmacology / HISTIDINA / HISTIDINE / hydro-lyases / Hydro-Lyases - antagonists & inhibitors / Hydro-Lyases - genetics / Hydro-Lyases - metabolism / IMMUNOELECTROPHORESE / immunoelectrophoresis / INHIBICION / INHIBITION / INMUNOELECTROFORESIS / isolation & purification / Kinetics / LIASAS / LYASE / MANGANESE / MANGANESO / Metabolism / Molecular Sequence Data / Organophosphonates / Organophosphonates - pharmacology / Parasitic plants. Weeds / PEPTIDE / peptides / PEPTIDOS / pharmacology / Phosphates / Phytopathology. Animal pests. Plant and forest protection / Plant cells / Plant physiology and development / Plants / polyacrylamide gel electrophoresis / Proteins / PROTEOLISIS / PROTEOLYSE / proteolysis / PURIFICACION / PURIFICATION / recombinant DNA / Recombinant Proteins / Recombinant Proteins - antagonists & inhibitors / Recombinant Proteins - genetics / Recombinant Proteins - isolation & purification / Spodoptera / SPODOPTERA FRUGIPERDA / transfection / Triazines / Triazoles / Triazoles - pharmacology / TRICHOPLUSIA NI / Triticum / Triticum - enzymology / Triticum - genetics / TRITICUM AESTIVUM / Weeds / Wheat