Fig. 3.

PP2C binds specifically to mGluR3 and not to other mGluRs. (A) The domains of mGluR1-8 used as bait in a yeast two-hybrid analysis are indicated on the left (amino acid residue numbers and the total length of each mGluR fragment are indicated). On the right, double transformants were spotted and grown on -LW or -LWH medium (see Fig. 1). (B) GST-mGluR3-830-879 or GST-mGluR2-821-872 were expressed in E. coli and purified by affinity chromatography; proteins were analyzed by SDS/PAGE and staining with Coomassie blue. (C) 6His-PP2Cα (1 μg) was incubated with 10 μg of immobilized GST-mGluR2-821-872 (GST-mGluR2) or GST-mGluR3-830-879 (GST-mGluR3). Retained proteins were analyzed by SDS/PAGE and immunoblotting with an anti-6His antibody.