FIGURE 16.
Mutation of tyrosine at position 184 as a part of the CRAC motif impacts membrane localization and transport activity of Oatp1d1. A, transport kinetics of Oatp1d1-Y184A in comparison with Oatp1d1-WT shows significant reduction in transport activity from Vm of 43.5 nmol/mg protein/min (c.i. 29.9–57.1) for Oatp1d1-WT to Vm of 12.14 nmol/mg protein/min (c.i. 8.8–15.5) for Oatp1d1-Y184A. B, partly impaired membrane localization of the Y184A mutant obtained by immunofluorescence analysis and Western blot stained with anti-His antibody shows a similar protein expression pattern between Oatp1d1-WT and Oatp1d1-Y184A cells. Immunocytochemistry was performed with FITC antibody (green) with nuclei stained with DAPI (blue), and samples were visualized on a confocal microscope.