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（Apicomplexa）胞内寄生虫依赖从宿主获取必需氨基酸。我们此前鉴定出一类顶复门特异性质膜定位氨基酸转运蛋白家族（ApiATs），并证实刚地弓形虫（Toxoplasma gondii）转运蛋白TgApiAT1可选择性摄取精氨酸。TgApiAT1对寄生虫的毒力至关重要，但在高浓度精氨酸的培养基中对寄生虫生长并非必需，这表明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。通过寄生虫实验结合非洲爪蟾（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，既可高亲和力摄取赖氨酸，也可低亲和力摄取精氨酸。TgApiAT6-1是致病阶段速殖子中主要的赖氨酸转运蛋白，对寄生虫增殖不可或缺。

本研究提供了一组液相色谱-质谱联用（LC-MS）数据集，对应在精氨酸或赖氨酸存在下表达TgApiAT6-1转运蛋白，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样本。该数据集检测了长达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，本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排的相关样本。该数据集附带已发表论文中图6对应的.raw原始数据文件：
《刚地弓形虫胞内寄生时多种转运蛋白协同获取必需阳离子氨基酸》
作者：Stephen J. Fairweather, Esther Rajendran, Martin Blume, Kiran Javed, Birte Steinhöfel, Malcolm J. McConville, Kiaran Kirk, Stefan Bröer, Giel G. van Dooren
预印本平台：bioRxiv 2021.06.25.450001; doi: https://doi.org/10.1101/2021.06.25.450001

本数据集为第三次重复液相色谱-质谱联用实验的数据，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（TIC）.raw输出文件。如需从每份.raw文件转换得到的已处理.mzML格式文件，可联系本文通讯作者。由于文件大小限制，本次未直接提供该类文件。本研究采用原始峰高对代谢物进行定量。卵母细胞胞内精氨酸与赖氨酸浓度通过引用论文中图S9的已知精氨酸/赖氨酸标准品的液相色谱-质谱校准曲线进行定量。