Cytosolic Action of Phytochelatin Synthase

• Published in: Plant physiology (Bethesda) 2010-05, Vol.153 (1), p.159-169
• Main Authors: Blum, Ralph, Meyer, Katrin C, Wünschmann, Jana, Lendzian, Klaus J, Grill, Erwin
• Format: Article
• Language: English
• Subjects: Aminoacyltransferases - genetics; Aminoacyltransferases - metabolism; Arabidopsis - enzymology; Arabidopsis - genetics; Arabidopsis Proteins - genetics; Arabidopsis Proteins - immunology; Azides; Azides - metabolism; Biological and medical sciences; Biosynthesis; Catabolism; Cytosol; Cytosol - enzymology; ENVIRONMENTAL STRESS AND ADAPTATION TO STRESS; Fundamental and applied biological sciences. Psychology; gamma-Glutamyltransferase - metabolism; Genetic Complementation Test; Glucosinolates; Glutathione - metabolism; Plant physiology and development; Plants; Protoplasts; Seedlings; Vacuoles; Vacuoles - metabolism; Xenobiotics
• ISSN: 0032-0889; 1532-2548; 1532-2548
• DOI: 10.1104/pp.109.149922

Glutathionylation of compounds is an important reaction in the detoxification of electrophilic xenobiotics and in the biosynthesis of endogenous molecules. The glutathione conjugates (GS conjugates) are further processed by peptidic cleavage reactions. In animals and plants, γ-glutamyl transpeptidases initiate the turnover by removal of the glutamate residue from the conjugate. Plants have a second route leading to the formation of γ-glutamylcysteinyl (γ-GluCys) conjugates. Phytochelatin synthase (PCS) is well known to mediate the synthesis of heavy metal-binding phytochelatins. In addition, the enzyme is also able to catabolize GS conjugates to the γ-GluCys derivative. In this study, we addressed the cellular compartmentalization of PCS and its role in the plant-specific γ-GluCys conjugate pathway in Arabidopsis (Arabidopsis thaliana). Localization studies of both Arabidopsis PCS revealed a ubiquitous presence of AtPCS1 in Arabidopsis seedlings, while AtPCS2 was only detected in the root tip. A functional AtPCS1:eGFP (enhanced green fluorescent protein) fusion protein was localized to the cytosolic compartment. Inhibition of the vacuolar import of GS-bimane conjugate via azide treatment resulted in both a strong accumulation of γ-GluCys-bimane and a massive increase of the cellular cysteine to GS-bimane ratio, which was not observed in PCS-deficient lines. These findings support a cytosolic action of PCS. Analysis of a triple mutant deficient in both Arabidopsis PCS and vacuolar γ-glutamyl transpeptidase GGT4 is consistent with earlier observations of an efficient sequestration of GS conjugates into the vacuole and the requirement of GGT4 for their turnover. Hence, PCS contributes specifically to the cytosolic turnover of GS conjugates, and AtPCS1 plays the prominent role. We discuss a potential function of PCS in the cytosolic turnover of GS conjugates.