Fig. 6

LmABCB3 is required for mitochondrial heme biosynthesis. a LmABCB3 cell levels correlate with the amount of heme synthesized from exogenous PPIX. The mitochondrial synthesis of heme from its precursor PPIX was measured as described in Experimental procedures after incubation of control, LmABCB3 ^+/− and LmABCB3 overexpressing L. major promastigotes with 0.5 μM PPIX. The results represent the mean ± SEM of three independent experiments. * p<0.05, ** p<0.02, *** p<0.001. b LmABCB3 interacts with heme. Solubilized membrane proteins of L. major parasites overexpressing LmABCB3-GFP were subjected to a pull-down assay with hemin-agarose in the absence (0) or the presence of the indicated increasing concentration of free hemin (upper panel) or free PPIX (lower panel). LmABCB3-GFP was detected by inmmunoblotting with Anti-GFP antibody (1:5000)