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)

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).

顶复门（Phylum Apicomplexa）胞内寄生虫依赖从宿主摄取必需氨基酸。我们此前鉴定出一类顶复门特异性质膜定位的氨基酸转运蛋白大家族——ApiAT转运蛋白（ApiATs），并证实刚地弓形虫（Toxoplasma gondii）转运蛋白TgApiAT1可介导精氨酸的选择性摄取。TgApiAT1对寄生虫毒力至关重要，但在精氨酸浓度较高的培养基中对寄生虫生长非必需，这提示至少存在另一种精氨酸转运蛋白。本研究鉴定出TgApiAT6-1为第二种精氨酸转运蛋白。通过结合寄生虫功能实验与非洲爪蟾（Xenopus laevis）卵母细胞中TgApiAT6-1的异源表达表征，我们证实TgApiAT6-1是一种广谱阳离子氨基酸转运蛋白，既可介导赖氨酸的高亲和力摄取，也可介导精氨酸的低亲和力摄取。TgApiAT6-1是致病型刚地弓形虫速殖子阶段的主要赖氨酸转运蛋白，对寄生虫增殖不可或缺。
本研究附带一份液相色谱-质谱联用法（Liquid Chromatography-Mass Spectrometry, LC-MS）数据集，对应两类样本：在精氨酸或赖氨酸存在下表达TgApiAT6-1转运蛋白的非洲爪蟾卵母细胞，以及在精氨酸存在下表达TgApiAT1转运蛋白的非洲爪蟾卵母细胞。该数据集测量了长达54小时时间进程中卵母细胞胞内代谢的变化，不仅追踪了转运蛋白对赖氨酸或精氨酸的摄取过程，还记录了卵母细胞内源性代谢物水平的波动。此外，数据集还包含在移除精氨酸或赖氨酸孵育液后30分钟、3小时及12小时时，已加载底物外流情况的相关样本。
本数据集为论文《Coordinated Action of Multiple Transporters in the Acquisition of Essential Cationic Amino Acids by the Intracellular Parasite Toxoplasma gondii》（作者：Stephen J. Fairweather、Esther Rajendran、Martin Blume、Kiran Javed、Birte Steinhöfel、Malcolm J. McConville、Kiaran Kirk、Stefan Bröer、Giel G. van Dooren，预印本发布于bioRxiv，编号2021.06.25.450001；DOI：https://doi.org/10.1101/2021.06.25.450001）中图6的配套原始数据文件。
本数据集为第三次重复LC-MS实验的数据，具体包含：
1. 样本汇总表（Excel文件），其中列有与上传的原始数据文件对应的编号样本ID；
2. 一个压缩（ZIP）文件夹，内含与汇总表中样本ID编号一致的单份原始数据文件，该文件夹包含样本汇总表中所有样本的总离子色谱图（Total Ion Chromatograms, TIC）.raw输出文件。如需从每份.raw文件转换得到的处理后.mzML格式文件，可联系本文通讯作者申请；因文件大小限制，本次未直接提供该类文件。
代谢物定量采用原始峰高法。卵母细胞内精氨酸与赖氨酸的浓度通过上述引用论文中图S9所示的已知精氨酸/赖氨酸标准品校准曲线进行定量。