Glycosylation Analysis of Feline Small Intestine Following Toxoplasma gondii Infection- raw datas 1

Toxoplasma gondii (T. gondii) is responsible for severe human and livestock diseases, huge economic losses, and adversely affects the health of the public and the development of animal husbandry. Glycosylation is a common posttranslational modification of proteins in eukaryotes, and N-glycosylation is closely related to the biological functions of proteins. However, glycosylation alterations in the feline small intestine following T. gondii infection have not been reported. In this study, the experimental group was intragastrically challenged with 600 brain cysts of the PRU strain that were collected from the infected mice. The cat's intestinal epithelial tissue was harvested at 10 days post-infection (DPI), and then sent for protein glycosylation analysis. High-performance liquid and mass spectrometry were used to analyze glycosylation alterations in the small intestine of cats infected with T. gondii. The results of the present study showed that 56 glycosylated peptides were upregulated and 37 glycosylated peptides were downregulated in the feline small intestine infected by T. gondii. Additionally, we also identified 8 N-glycosylated proteins of T. gondii, including 8 N-glycopeptides and 8 N-glycosylation sites. The protein A0A086JND6_TOXGO (eEF2) and its corresponding peptide sequence were identified in T. gondii infection. During the process of glycosylation of T. gondii oocysts, some special GO terms (cellular process and metabolic process, cell and cell part, catalytic activity) were significantly enriched, and the COG function prediction results showed that posttranslational modification, protein turnover, and chaperones (11%) had the highest enrichment for T. gondii. Interestingly, eEF2 is also involved in the significantly enriched T. gondii MAPK pathway. Protein–protein interaction networks were analyzed by MCODE_Cluster of Cytoscaype for the differentially expressed peptides/proteins. The host proteins ICAM-1 and PPT1 and the endoplasmic reticulum stress pathway may play an important role in the glycosylation of Toxoplasma-infected hosts. This is the first report showing that T. gondii oocysts can undergo N-glycosylation and eEF2 is involved, which provides a new target for anti-T. gondii therapy to prevent the spread of T. gondii oocysts in the future.

刚地弓形虫（T. gondii）是导致人类及家畜严重疾病、巨大经济损失的病原体，严重影响了公众健康与畜牧业的发展。糖基化是真核生物中蛋白质常见的翻译后修饰，而N-糖基化与蛋白质的生物学功能密切相关。然而，关于刚地弓形虫感染后猫的小肠中糖基化改变尚未有报道。在本研究中，实验组通过胃内投喂600个由感染小鼠收集的PRU菌株脑囊虫进行挑战。感染后10天（DPI）收获猫的肠上皮组织，并进行蛋白质糖基化分析。采用高效液相色谱和质谱技术分析感染刚地弓形虫的猫的小肠中糖基化改变。本研究结果显示，在感染刚地弓形虫的猫的小肠中，共有56种糖基化肽被上调表达，而37种糖基化肽被下调表达。此外，我们还鉴定了8种刚地弓形虫的N-糖基化蛋白，包括8种N-糖肽和8个N-糖基化位点。在刚地弓形虫感染过程中，蛋白质A0A086JND6_TOXGO（eEF2）及其相应的肽序列被鉴定。在刚地弓形虫子囊的糖基化过程中，一些特殊的GO术语（细胞过程和代谢过程、细胞和细胞部分、催化活性）显著富集，COG功能预测结果显示，翻译后修饰、蛋白质周转和伴侣蛋白（11%）在刚地弓形虫中富集度最高。有趣的是，eEF2还参与了显著富集的刚地弓形虫MAPK途径。通过MCODE聚类方法分析了差异表达肽/蛋白的蛋白质-蛋白质相互作用网络。宿主蛋白ICAM-1和PPT1以及内质网应激途径可能在感染弓形虫宿主的糖基化过程中发挥重要作用。这是首次报道刚地弓形虫子囊可以发生N-糖基化，且eEF2参与其中，这为未来预防刚地弓形虫子囊传播的抗刚地弓形虫治疗提供了新的靶点。