Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, São Paulo, Brazil.
Departamento de Química Analítica, Instituto de Química, Universidade Federal da Bahia, Salvador, BA, Brazil.
PLoS Negl Trop Dis. 2019 Feb 6;13(2):e0007103. doi: 10.1371/journal.pntd.0007103. eCollection 2019 Feb.
Trypanosoma cruzi, the etiological agent of Chagas' disease, affects 8 million people predominantly living in socioeconomic underdeveloped areas. T. cruzi trypomastigotes (Ty), the classical infective stage, interact with the extracellular matrix (ECM), an obligatory step before invasion of almost all mammalian cells in different tissues. Here we have characterized the proteome and phosphoproteome of T. cruzi trypomastigotes upon interaction with ECM (MTy) and the data are available via ProteomeXchange with identifier PXD010970. Proteins involved with metabolic processes (such as the glycolytic pathway), kinases, flagellum and microtubule related proteins, transport-associated proteins and RNA/DNA binding elements are highly represented in the pool of proteins modified by phosphorylation. Further, important metabolic switches triggered by this interaction with ECM were indicated by decreases in the phosphorylation of hexokinase, phosphofructokinase, fructose-2,6-bisphosphatase, phosphoglucomutase, phosphoglycerate kinase in MTy. Concomitantly, a decrease in the pyruvate and lactate and an increase of glucose and succinate contents were detected by GC-MS. These observations led us to focus on the changes in the glycolytic pathway upon binding of the parasite to the ECM. Inhibition of hexokinase, pyruvate kinase and lactate dehydrogenase activities in MTy were observed and this correlated with the phosphorylation levels of the respective enzymes. Putative kinases involved in protein phosphorylation altered upon parasite incubation with ECM were suggested by in silico analysis. Taken together, our results show that in addition to cytoskeletal changes and protease activation, a reprogramming of the trypomastigote metabolism is triggered by the interaction of the parasite with the ECM prior to cell invasion and differentiation into amastigotes, the multiplicative intracellular stage of T. cruzi in the vertebrate host.
克氏锥虫，恰加斯病的病原体，主要影响生活在社会经济欠发达地区的 800 万人。克氏锥虫的锥虫（Ty），经典的感染阶段，与细胞外基质（ECM）相互作用，这是在不同组织的几乎所有哺乳动物细胞入侵之前的一个必要步骤。在这里，我们已经描述了克氏锥虫锥虫与细胞外基质（MTy）相互作用时的蛋白质组和磷酸蛋白质组，数据可通过 ProteomeXchange 以标识符 PXD010970 获得。参与代谢过程（如糖酵解途径）、激酶、鞭毛和微管相关蛋白、运输相关蛋白和 RNA/DNA 结合元件的蛋白质在磷酸化修饰的蛋白质池中高度代表。此外，由与 ECM 的这种相互作用触发的重要代谢开关通过 MTy 中己糖激酶、磷酸果糖激酶、果糖-2,6-二磷酸酶、磷酸葡糖变位酶、磷酸甘油酸激酶的磷酸化减少来指示。同时，通过 GC-MS 检测到丙酮酸和乳酸的减少以及葡萄糖和琥珀酸含量的增加。这些观察结果使我们专注于寄生虫与 ECM 结合后糖酵解途径的变化。在 MTy 中观察到己糖激酶、丙酮酸激酶和乳酸脱氢酶活性的抑制，并且这与相应酶的磷酸化水平相关。通过计算机分析建议了参与 ECM 孵育后蛋白质磷酸化改变的假定激酶。总之，我们的结果表明，除了细胞骨架变化和蛋白酶激活之外，寄生虫与 ECM 相互作用还会触发锥虫代谢的重编程，然后再入侵和分化为阿米巴虫，这是克氏锥虫在脊椎动物宿主中的增殖性细胞内阶段。
