DOI: 10.17632/4xy7zppt2s.1
title: Coordinated Action of Multiple Transporters in the Acquisition of Essential Cationic Amino Acids by the Intracellular Parasite Toxoplasma gondii (LC-MS Repeat Data set 3)

description: Intracellular parasites of the phylum Apicomplexa are dependent on the scavenging of essential amino acids from their hosts. We previously identified a large family of apicomplexan-specific plasma membrane-localized amino acid transporters, the ApiATs, and showed that the Toxoplasma gondii transporter TgApiAT1 functions in the selective uptake of arginine. TgApiAT1 is essential for parasite virulence, but dispensable for parasite growth in medium containing high concentrations of arginine, indicating the presence of at least one other arginine transporter. Here we identify TgApiAT6-1 as the second arginine transporter. Using a combination of parasite assays and heterologous characterisation of TgApiAT6-1 in Xenopus laevis oocytes, we demonstrate that TgApiAT6-1 is a general cationic amino acid transporter that mediates both the high-affinity uptake of lysine and the low-affinity uptake of arginine. TgApiAT6-1 is the primary lysine transporter in the disease-causing tachyzoite stage of T. gondii and is essential for parasite proliferation. Here we provide an LC-MS data set of Xenopus laevis oocytes expressing the TgApiAT6-1 transporter in the presence of Arg or Lys and the TgApiAT1 transporter in the presence of Arg. This data set measures the change in intracellular oocyte metabolism over a time course of up to 54 hours, charting the uptake of Lys or Arg by the transporters in addition to fluctuations in endogenous oocyte metabolite levels. In addition, samples are also present that demonstrate the efflux of the loaded substrates at 30min, 3hr, and 12hr following the removal of either the Arg or Lys incubating solution. This data set provides the .raw data files which accompanies Fig. 6 in the publication: Coordinated Action of Multiple Transporters in the Acquisition of Essential Cationic Amino Acids by the Intracellular Parasite Toxoplasma gondii Stephen J. Fairweather, Esther Rajendran, Martin Blume, Kiran Javed, Birte Steinhöfel, Malcolm J. McConville, Kiaran Kirk, Stefan Bröer, Giel G. van Dooren bioRxiv 2021.06.25.450001; doi: https://doi.org/10.1101/2021.06.25.450001 This is the data set for the 3rd repeat LC-MS experiment, containing: 1. Sample Summary Table (Excel file) of including samples IDs (numbered) corresponding to the uploaded raw data files. 2. A compressed (zipped) folder containing individual raw data files numbered with the same sample ID as in the summary spreadsheet. The folder contains a .raw output file of the Total Ion Chromatograms (TIC) from each sample in the Sample Summary Table. Processed .mzml files converted form each sample .raw file can be provided upon request of the authors from the publication above. They are not provided directly here due to file size constraints. Raw peak height was used for the quantification of metabolites. Intracellular Arg and Lys oocyte concentrations were quantified using the LC-MS calibration curves with known Arg/Lys standards (in Fig. S9 of the cited publication).