Molecular characterization of the homo-phytochelatin synthase of soybean Glycine max: relation to phytochelatin synthase.
The phytochelatin homologs homo-phytochelatins are heavy metal-binding peptides present in many legumes. To study the biosynthesis of these compounds, we have isolated and functionally expressed a cDNA GmhPCS1 encoding homo-phytochelatin synthase from Glycine max, a plant known to accumulate homo-phytochelatins rather than phytochelatins upon the exposure to heavy metals. The catalytic properties of GmhPCS1 were compared with the phytochelatin synthase AtPCS1 from Arabidopsis thaliana. When assayed only in the presence of glutathione, both enzymes catalyzed phytochelatin formation. GmhPCS1 accepted homoglutathione as the sole substrate for the synthesis of homo-phytochelatins whereas AtPCS1 did not. Homo-phytochelatin synthesis activity of both recombinant enzymes was significantly higher when glutathione was included in the reaction mixture. The incorporation of both glutathione and homoglutathione into homo-phytochelatin, n = 2, was demonstrated using GmhPCS1 and AtPCS1. In addition to bis(glutathionato)-metal complexes, various other metal-thiolates were shown to contribute to the activation of phytochelatin synthase. These complexes were not accepted as substrates by the enzyme, thereby suggesting that a recently proposed model of activation cannot fully explain the catalytic mechanism of phytochelatin synthase (Vatamaniuk, O. K., Mari, S., Lu, Y. P., and Rea, P. A. (2000) J. Biol. Chem. 275, 31451-31459).
Oven
Matjaz
M
Leibniz-Institut für Pflanzenbiochemie, Weinberg 3 and the Biozentrum der Universität Halle, Weinbergweg 22, Halle/Saale 06120, Germany.
Page
Jonathan E
JE
Zenk
Meinhart H
MH
Kutchan
Toni M
TM
D000595:Amino Acid Sequence; D019881:Aminoacyltransferases / Q000737:chemistry* / Q000235:genetics*; D017360:Arabidopsis / Q000201:enzymology; D001665:Binding Sites; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D013058:Mass Spectrometry; D008670:Metals / Q000378:metabolism; D008956:Models, Chemical; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013025:Glycine max / Q000201:enzymology*; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013997:Time Factors
D018076:DNA, Complementary; D008670:Metals; D011994:Recombinant Proteins; D013438:Sulfhydryl Compounds; D019881:Aminoacyltransferases; C093784:glutathione gamma-glutamylcysteinyltransferase