title: Abscisic acid binds to recombinant Arabidopsis thaliana G-protein coupled receptor-type G-protein 1 in Sacaromycese cerevisiae and in vitro.
abstract: The G-protein coupled receptor-type G-proteins (GTG) 1 and 2 from Arabidopsis thaliana have been proposed to function in the modulation of abscisic acid (ABA) mediated responses to stress and development. In particular it has been suggested that they function as ABA receptors based on in planta and in vitro analyses. However a recent independent report was inconsistent with this, suggesting that there is no link between the GTGs and ABA in planta. Here we provide an independent assessment of the ability of ABA to bind to recombinant GTG1 in vitro and in vivo in Sacaromycese cerevisiae. Radio-labelled binding assays on enriched lipid-reconstituted recombinant GTG1, demonstrated specific concentration dependent binding of [(3)H]-ABA with a dissociation constant (KD) of 80 nM, corroborating previous reports. Assessment of the binding of [(3)H]-ABA to intact GTG1 expressing yeast, showed GTG1-dependent binding in vivo, yielding a physiologically relevant KD of 0.6 μM. Together these results provide independent evidence of a binding-interaction between ABA and GTG1 in vitro and in vivo, in support of the previously proposed possibility of a biologically relevant interaction between GTG1 and ABA.
authors.affiliation: National Research Council of Canada, 110 Gymnasium Place, Saskatoon, SK S7N 0W9, Canada.
authors.forename: Olesya A
authors.initials: OA
authors.lastname: Kharenko
authors.forename: Pooja
authors.initials: P
authors.lastname: Choudhary
authors.forename: Michele C
authors.initials: MC
authors.lastname: Loewen
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D043562:Receptors, G-Protein-Coupled / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D043562:Receptors, G-Protein-Coupled; D011994:Recombinant Proteins; D000040:Abscisic Acid; C534590:GTG1 protein, Arabidopsis