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. Author manuscript; available in PMC: 2011 Sep 1.

Published in final edited form as: Biochim Biophys Acta. 2010 May 28;1798(9):1757–1766. doi: 10.1016/j.bbamem.2010.05.017

Fig. 5 — A). (i) Photoaffinity labeling of WT-CaCdr1p and its mutant variants with [¹²⁵I]-IAAP. The PM fraction (30 μg protein) of cells expressing WT-CaCdr1p and its mutant variants were incubated with 7.5 nM [¹²⁵I]-IAAP (2300 Ci/mmol). The samples were UV crosslinked and processed as described elsewhere [34]. (ii) Western blot analysis using anti-GFP antibody to ensure equal loading of WT-CaCdr1p and its Signature mutant variants (iii) Normalized incorporated [¹²⁵I] IAAP labeling with Western blot intensity. The values are shown in percentage. B). Photoaffinity labeling of WT-CaCdr1p and its mutant variants with [α-³²P] 8-azido ATP. The PM fraction (30 μg) of cells expressing the WT-CaCdr1p and its mutant variants were incubated with 10 μM [α-³²P] 8-azido ATP 7.5 μCi/nmol at 4°C and competed with 10 mM cold ATP (+ATP lane) as described in [34]. (ii) Western blot analysis using anti-GFP antibody to ensure equal loading of WT-CaCdr1p and its Signature mutant variants (iii) Normalized incorporated [α-³²P] 8-azido ATP labeling with Western blot intensity. The values are shown in percentage.

Fig. 5 — A). (i) Photoaffinity labeling of WT-CaCdr1p and its mutant variants with [¹²⁵I]-IAAP. The PM fraction (30 μg protein) of cells expressing WT-CaCdr1p and its mutant variants were incubated with 7.5 nM [¹²⁵I]-IAAP (2300 Ci/mmol). The samples were UV crosslinked and processed as described elsewhere [34]. (ii) Western blot analysis using anti-GFP antibody to ensure equal loading of WT-CaCdr1p and its Signature mutant variants (iii) Normalized incorporated [¹²⁵I] IAAP labeling with Western blot intensity. The values are shown in percentage. B). Photoaffinity labeling of WT-CaCdr1p and its mutant variants with [α-³²P] 8-azido ATP. The PM fraction (30 μg) of cells expressing the WT-CaCdr1p and its mutant variants were incubated with 10 μM [α-³²P] 8-azido ATP 7.5 μCi/nmol at 4°C and competed with 10 mM cold ATP (+ATP lane) as described in [34]. (ii) Western blot analysis using anti-GFP antibody to ensure equal loading of WT-CaCdr1p and its Signature mutant variants (iii) Normalized incorporated [α-³²P] 8-azido ATP labeling with Western blot intensity. The values are shown in percentage.