Fig. 1.

UT-A1 urea transporter associates with dynamin. A: UT-A1-HEK293 cells were transfected with pEGFP, pEGFP-dynamin, or pEGFP-dynamin K44A for 48 h. Cells were lysed with RIPA buffer and the supernatants were used for immunoprecipitation with UT-A1 antibody. Total cell lysates or UT-A1-precipitated samples were processed for Western blot with GFP antibody. B: dynamin binds to UT-A1 intracellular loop. EGFP-Dynamin and EGFP-K44A were prepared by transfection of pEGFP-Dyn or pEGFP-K44A into HEK293 cells. The proteins were precipitated with GFP antibody and conjugated to the protein A beads. An equal amount of [³⁵S]-labeled N-, C-, and Lp-A1 synthesized in reticulocyte lysates was added to GFP-Dyn or GFP-K44A beads for pulldown assay. The three fragments of N-, C-, and Lp-A1 tagged with c-myc were precipitated with c-myc antibody as control. The [³⁵S] signal was detected by autoradiography.