title: The Arabidopsis phi class glutathione transferase AtGSTF2: binding and regulation by biologically active heterocyclic ligands.
abstract: The plant-specific phi class of glutathione transferases (GSTFs) are often highly stress-inducible and expressed in a tissue-specific manner, suggestive of them having important protective roles. To date, these functions remain largely unknown, although activities associated with the binding and transport of reactive metabolites have been proposed. Using a sensitive and selective binding screen, we have probed the Arabidopsis thaliana GSTFs for natural product ligands from bacteria and plants. Uniquely, when overexpressed in bacteria, family members GSTF2 and GSTF3 bound a series of heterocyclic compounds, including lumichrome, harmane, norharmane and indole-3-aldehyde. When screened against total metabolite extracts from A. thaliana, GSTF2 also selectively bound the indole-derived phytoalexin camalexin, as well as the flavonol quercetin-3-O-rhamnoside. In each case, isothermal titration calorimetry revealed high-affinity binding (typically Kd<1 μM), which was enhanced in the presence of glutathione and by the other heterocyclic ligands. With GSTF2, these secondary ligand associations resulted in an allosteric enhancement in glutathione-conjugating activity. Together with the known stress responsiveness of GSTF2 and its association with membrane vesicles, these results are suggestive of roles in regulating the binding and transport of defence-related compounds in planta.
authors.affiliation: Centre for Bioactive Chemistry, School of Biological and Biomedical Sciences, Durham University, South Road, Durham DH1 3LE, UK.
authors.forename: David P
authors.initials: DP
authors.lastname: Dixon
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Sellars
authors.forename: Robert
authors.initials: R
authors.lastname: Edwards
mesh_terms: D000890:Anti-Infective Agents / Q000494:pharmacology; D017360:Arabidopsis / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D006571:Heterocyclic Compounds / Q000737:chemistry* / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology; D000094162:Phytoalexins
chemical_list: D000890:Anti-Infective Agents; D029681:Arabidopsis Proteins; D005982:Glutathione Transferase; D006571:Heterocyclic Compounds; D007211:Indoles; D011993:Recombinant Fusion Proteins; D013844:Thiazoles; C102405:camalexin; D000094162:Phytoalexins; C478066:GSTF2 protein, Arabidopsis