Skip to main content
. 2009 Jan 16;284(3):1820–1830. doi: 10.1074/jbc.M804590200

FIGURE 2.

FIGURE 2.

PMCA1 binding to NHERF-2 in HT-29 cells is mediated by the C-terminal PDZ motif. A, GST pull-downs using pan-PMCA antibody showing that PMCA interacted with NHERF-2 but not NHERF-1 in HT-29 cells, and the interaction was mediated by the second PDZ module (PDZ2) of NHERF-2. B, co-immunoprecipitations investigating the interaction between NHERF1/2 and PMCA. Lane a, HT-29 cell lysate control; lanes b and c, immunoprecipitation with anti-NHERF-1 antibody; lanes d and e, immunoprecipitation with anti-NHERF-2 antibody. B, panel i, blot probed with pan-PMCA antibody. B, panel ii, blot probed with anti-PMCA4 antibody. NHERF-2 interacted with PMCA, and this interaction occurred in the presence and absence of ACh. C, Western blot showing HEK293 cells transfected with GFP-tagged PMCA1b C terminus ± PDZ binding motif expressed proteins of the predicted size (∼50 kDa). D, pull-downs using GFP constructs of the C terminus of PMCA1b ± PDZ binding motif together with GST-NHERF-2. The interaction with NHERF-2 was abolished when the C-terminal ETSL motif was deleted (GFP-PMCA1b-DPDZ). Western blots are representative of at least three individual experiments.