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. 2008 Dec 12;283(50):35010–35022. doi: 10.1074/jbc.M806446200

FIGURE 5.

FIGURE 5.

Interaction of WT and mutant Pdr5 with the drug substrate analogue [125I]IAAP. A, photoaffinity cross-linking of Pdr5 to [125I]IAAP was carried out in purified PM vesicles as described under “Experimental Procedures.” Quantification of the 125I signal in the Pdr5 band in plasma membranes prepared from the Δpdr5 strain AD1–7, the double-copy WT- yeast strain JG2004, and the double-copy S558Y mutant strain (JG 2010) was performed with a STORM 860 PhosphorImager system. Cross-linking of [125I]IAAP was carried out in the absence (filled bars) or presence (empty bars) of 100 mm clotrimazole. B, photoaffinity cross-linking of [125I]IAAP to WT (•) or S558Y (□) Pdr5 was carried out as described in A in the presence of increasing concentrations of clotrimazole. The 125I signal in the Pdr5 band was quantified and plotted with the curve-fitting GraphPad Prism software. The inset depicts the data with an expanded x axis in the range 0–100 μm ATP. The x and y axis legends for the inset are the same as those for the main figure. Representative plots are shown (n = 2).