The Inositol Pyrophosphate Biosynthetic Pathway of Trypanosoma cruzi,ACS Chemical Biology

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The Inositol Pyrophosphate Biosynthetic Pathway of Trypanosoma cruzi
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2021-01-07 , DOI: 10.1021/acschembio.0c00759
Brian S Mantilla ₁ , Leticia D Do Amaral ₁ , Henning J Jessen ₂ , Roberto Docampo ₁

Affiliation

Inositol phosphates (IPs) are phosphorylated derivatives of myo-inositol involved in the regulation of several cellular processes through their interaction with specific proteins. Their synthesis relies on the activity of specific kinases that use ATP as phosphate donor. Here, we combined reverse genetics and liquid chromatography coupled to mass spectrometry (LC-MS) to dissect the inositol phosphate biosynthetic pathway and its metabolic intermediates in the main life cycle stages (epimastigotes, cell-derived trypomastigotes, and amastigotes) of Trypanosoma cruzi, the etiologic agent of Chagas disease. We found evidence of the presence of highly phosphorylated IPs, like inositol hexakisphosphate (IP₆), inositol heptakisphosphate (IP₇), and inositol octakisphosphate (IP₈), that were not detected before by HPLC analyses of the products of radiolabeled exogenous inositol. The kinases involved in their synthesis (inositol polyphosphate multikinase (TcIPMK), inositol 5-phosphate kinase (TcIP5K), and inositol 6-phosphate kinase (TcIP6K)) were also identified. TcIPMK is dispensable in epimastigotes, important for the synthesis of polyphosphate, and critical for the virulence of the infective stages. TcIP5K is essential for normal epimastigote growth, while TcIP6K mutants displayed defects in epimastigote motility and growth. Our results demonstrate the relevance of highly phosphorylated IPs in the life cycle of T. cruzi.

中文翻译：

克氏锥虫肌醇焦磷酸生物合成途径

磷酸肌醇 (IP) 是肌醇的磷酸化衍生物，通过与特定蛋白质的相互作用参与多种细胞过程的调节。它们的合成依赖于使用 ATP 作为磷酸盐供体的特定激酶的活性。在这里，我们将反向遗传学和液相色谱联用质谱（LC-MS）相结合，剖析了克氏锥虫主要生命周期阶段（表鞭毛体、细胞源性锥鞭毛体和无鞭毛体）的磷酸肌醇生物合成途径及其代谢中间体，恰加斯病的病原体。我们发现了高度磷酸化 IP 存在的证据，如肌醇六磷酸 (IP ₆ )、肌醇七磷酸 (IP ₇ ) 和肌醇八磷酸 (IP ₈ )，这些是以前通过对放射性标记的外源肌醇产物进行 HPLC 分析时未检测到的。还鉴定了参与其合成的激酶（肌醇多磷酸激酶 (TcIPMK)、肌醇 5-磷酸激酶 (TcIP5K) 和肌醇 6-磷酸激酶 (TcIP6K)）。TcIPMK在上鞭毛体中是可有可无的，对于多磷酸盐的合成很重要，并且对于感染阶段的毒力至关重要。TcIP5K对于正常的上鞭毛体生长至关重要，而TcIP6K突变体在上鞭毛体运动和生长方面表现出缺陷。我们的结果证明了高度磷酸化 IP 在T. cruzi生命周期中的相关性。

更新日期：2021-02-19

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