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. 2013 May 2;9(5):e1003331. doi: 10.1371/journal.ppat.1003331

Figure 1. Toxoplasma TgNST1 is a nucleotide-sugar transporter that transports UDP-GlcNAc and UDP-GalNac.

Figure 1

A) Analysis of protein expression by western blot. Total extracts from Saccharomyces cerevisiae strain PRY225 and the same strain transformed with the p426 vector encoding the Toxoplasma NST1HA protein (TgNST1) were resolved by SDS-PAGE followed by transfer to PVDF membrane. Detection of the recombinant protein was done using an anti-HA antibody. B) 1 mg of membrane vesicles from yeast transformed with an empty vector or the vector encoding the TgNST1HA protein were incubated with radioactive and unlabeled UDP-GlcNAc or UDP-GalNAc for 4 min. Final concentration of nucleotide-sugar is 2 µM per sample. Vesicles were separated by centrifugation and radioactivity within vesicles was measured in the pellet after acid precipitation. Transport activity is calculated as the difference of radioactive solutes in the vesicle pellet after incubation at 30°C vs 0°C and expressed as pmol of nucleotide sugar/mg protein/min.