Fig. 3: Unspecific uptake of rGLUT5, lipidomic analysis of lipid extracts, and their respective stabilization.

a Uptake of ¹⁴C-d-glucose by rGLUT5 in proteoliposomes and empty liposomes prepared using soya PC represented as empty circles and squares, respectively. Inset; uptake of rGLUT5 (empty bars) and empty liposomes (red bars) at one-time point (2 min) in soya PC for glucose and fructose. Data were normalized to the uptake of rGLUT5 as 100%, and axis labels were not shown for clarity. Error bars indicate mean ± s.e.m. of n = 3 independent experiments. b Relative 2 min uptake of ¹⁴C-d-glucose by WT rGLUT5 (empty bars) and A395W (black bars) reconstituted into liposomes made from soya PC lipids. Non-specific uptake (red bars) into liposomes without protein is also represented. Data were normalized to WT rGLUT5 transport activity; error bars indicate mean ± s.e.m. of n = 3 independent experiments. c Uptake of ¹⁴C-d-glucose by PfHT1 (Plasmodium falciparum Hexose Transporter) in proteliposomes prepared using brain-fraction-seven and soya PC extracts, respectively, in the absence (empty bars) and presence (black bars) of inhibitor MMV009085 at 30 s. Non-specific ¹⁴C-d-Glucose uptake (red bars) is represented as in (b), and data was normalized to PfHT1 transport activity; error bars indicate mean ± s.e.m. of n = 3 independent experiments. d Lipidomics analysis of crude extracts from Fig. 1a. Phosphatidylcholine (PC) in black; ceramide (Cer) in light yellow; phosphatidylethanolamine (PE) in light blue; sphingomyelin (SM) in dark green; phosphatidylserine (PS) in light gray; phosphatidylinositol (PI) in light orange; monogalactosyldiacylglycerol (MGDG) in red; diacylglyceryltrimethylhomo-Ser (DGTS) in pink; and triglycerides (TG) in light green. e Melting curves of rGLUT5 in the presence of individual phospholipids, as colored in (d). As a control, the melting temperature of rGLUT5 with DDM added, but no lipid is shown in red; the data are normalized fluorescence mean ± s.e.m. of n = 3 independent experiments. f The melting temperature for rGLUT5 in the presence of individual phospholipids, as calculated from the curves in panel (e). Error bars indicate mean ± s.e.m. of n = 3 independent experiments.