Univ. Lille, CNRS, Inserm, CHU Lille, Institut Pasteur de Lille, U1019-UMR 9017-CIIL-Centre d'Infection et d'Immunité de Lille, 59000 Lille, France.
Proteomics Platform Necker (PPN), Université de Paris, Structure Fédérative de Recherche Necker (SFR Necker, INSERM US24/CNRS UAR3633), 75015 Paris, France.
Int J Mol Sci. 2022 Jan 19;23(3):1069. doi: 10.3390/ijms23031069.
Malaria parasites require multiple phosphorylation and dephosphorylation steps to drive signaling pathways for proper differentiation and transformation. Several protein phosphatases, including protein phosphatase 1 (PP1), one of the main dephosphorylation enzymes, have been shown to be indispensable for the life cycle. The catalytic subunit of PP1 (PP1c) participates in cellular processes via dynamic interactions with a vast number of binding partners that contribute to its diversity of action. In this study, we used transgenic parasite strains stably expressing PP1c or its inhibitor 2 (I2) tagged with mCherry, combined with the mCherry affinity pulldown of proteins from asexual and sexual stages, followed by mass spectrometry analyses. Mapped proteins were used to identify interactomes and to cluster functionally related proteins. Our findings confirm previously known physical interactions of PP1c and reveal enrichment of common biological processes linked to cellular component assembly in both schizonts and gametocytes to biosynthetic processes/translation in schizonts and to protein transport exclusively in gametocytes. Further, our analysis of PP1c and I2 interactomes revealed that nuclear export mediator factor and peptidyl-prolyl cis-trans isomerase, suggested to be essential in , could be potential targets of the complex PP1c/I2 in both asexual and sexual stages. Our study emphasizes the adaptability of PP1 and provides a fundamental study of the protein interaction landscapes involved in a myriad of events in , suggesting why it is crucial to the parasite and a source for alternative therapeutic strategies.
疟原虫需要多次磷酸化和去磷酸化步骤来驱动信号通路，以实现适当的分化和转化。几种蛋白磷酸酶，包括蛋白磷酸酶 1（PP1），作为主要的去磷酸化酶之一，已被证明对生命周期是不可或缺的。PP1 的催化亚基（PP1c）通过与大量结合伴侣的动态相互作用参与细胞过程，这有助于其多样化的作用。在这项研究中，我们使用稳定表达 PP1c 或其抑制剂 2（I2）标记为 mCherry 的转基因寄生虫株，结合有丝分裂和有性阶段的 mCherry 亲和下拉法，随后进行质谱分析。映射的蛋白质用于鉴定相互作用组，并将功能相关的蛋白质聚类。我们的发现证实了 PP1c 的先前已知的物理相互作用，并揭示了在裂殖子和配子体中与细胞成分组装相关的常见生物学过程的富集，在裂殖子中与生物合成过程/翻译相关，而在配子体中仅与蛋白质运输相关。此外，我们对 PP1c 和 I2 相互作用组的分析表明，核输出介质因子和肽基脯氨酰顺反异构酶，被认为在 中是必不可少的，可能是复杂的 PP1c/I2 在无性和有性阶段的潜在靶点。我们的研究强调了 PP1 的适应性，并为涉及 的无数事件的蛋白质相互作用景观提供了基础研究，这表明为什么它对寄生虫至关重要，并且是替代治疗策略的来源。
