FIGURE 7.

Growth stage regulation of S-adenosylmethionine transport activity. Accumulation of 60 nm S-[³H]adenosylmethionine was measured along the growth phases (48, 72, 96, and 120 h) for 10 min in L. infantum wild type (white bars), LV39SNF4000.4 transfected with AdoMetT1-GFP (gray bars), and L. major LV39 wild type (black bars). Average of triplicate measures is shown. Western blot with an anti-GFP antibody or with a control α-tubulin antibody to monitor the amounts of proteins layered was carried out with LV39SNF4000.4 transfected with AdoMetT1-GFP as described under “Materials and Methods.” Cellular localization of AdoMetT1 was deduced from fluorescence microscopy analysis of LV39SNF4000.4 transfected with the AdoMetT1 gene fused in-frame with the GFP gene. The majority of the fluorescence derived from the AdoMetT1-GFP hybrid protein was at the level of the plasma membrane throughout the various growth stages.