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. Author manuscript; available in PMC: 2010 Mar 1.
Published in final edited form as: Biochim Biophys Acta. 2009 Apr 21;1788(9):1752–1761. doi: 10.1016/j.bbamem.2009.04.009

Figure 3.

Figure 3

Expression and drug binding assay using plasma membranes cells expressing WT CaCdr1p-GFP and the mutant variants of TMS 5 in S. cerevisiae. (A) Western analysis showing proper membrane localization of WT CaCdr1p-GFP and its mutant variants. (B) Photo affinity labeling of WT CaCdr1p-GFP and its mutant variants with [125I] IAAP in the presence and absence of NYS. The PM proteins (25μg) were photoaffinity labeled with [125I] IAAP under subdued light as described in Materials and Methods. Upper panel, lane 1, WT CaCdr1p-GFP (no drug); lane 2, WT CaCdr1p-GFP with NYS; lanes 3, 5, 7 show [125I] IAAP binding in mutant variants of CaCdr1p-GFP without drug while lanes 4, 6, 8 exhibit [125I] IAAP binding in the presence of 200μM NYS. Lower panel depicts Western analysis to show equal loading of CaCdr1p-GFP.