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. 2013 Jun 21;288(31):22658–22669. doi: 10.1074/jbc.M113.485284

FIGURE 5.

FIGURE 5.

Liposome-reconstituted ABCB6 shows ATP-dependent transport of molecules identified in a HTS assay. A, liposome reconstituted ABCB6 interacts with both tomatine and verteporfin. A, top panel, ABCB6 liposomes (4 μg of protein) were subjected to hemin affinity chromatography in the presence or absence of molecules identified in the HTS assay, and bound protein was subjected to 4–15% SDS-PAGE followed by Western blot using FLAG M2 antibody. Bottom panel, percent binding was estimated by plotting the band intensities corresponding to untreated ABCB6, which was set to 100%. B, liposome-reconstituted ABCB6 shows increased ATPase activity in the presence of molecules identified in the HTS assay. Values represent the mean ± S.D. *, significantly different from basal ATPase activity; p < 0.01. &, significantly different from coproporphyrinogen (Copro III) ATPase activity; p < 0.05. #, significantly different from verteporfin ATPase activity; p < 0.01. C and D, liposome-reconstituted ABCB6 shows ATP-dependent transport of verteporfin (C) and tomatine (D). Kinetic parameters were determined by linear regression analysis of the Lineweaver-Burk transformation of the data points.