A phosphoproteomic approach towards the understanding of the role of TGF-β in Trypanosoma cruzi biology.
A phosphoproteomic approach towards the understanding of the role of TGF-β in Trypanosoma cruzi biology.
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DOI：
10.1371/journal.pone.0038736
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Waghabi MC
中科院分区：
文献类型：
--
作者：
Ferrão PM;de Oliveira FL;Degrave WM;Araujo-Jorge TC;Mendonça-Lima L;Waghabi MC
转化生长因子β（Transforming growth factor beta，TGF-β）在恰加斯病中起着关键作用，不仅在恰加斯病心肌病的发展中，而且在恰加斯病的许多阶段都起着重要作用。cruzi生命周期和在宿主细胞环境中的存活。T. cruzi调节这些机制的作用仍然未知。为了鉴定参与TGF-β反应的寄生虫蛋白，我们利用二维凝胶电泳（2DE）分析和质谱（MS）蛋白鉴定的组合方法。通过TGF-β的信号传导依赖于磷酸化事件，磷酸化是调节基因表达的最相关和普遍存在的翻译后修饰之一，特别是在锥虫中，因为它们缺乏几种转录控制机制。在这里，我们展示了一个动力学的观点，T。TGF-β作用1、5、30和60 min后，可促进寄生虫磷酸化网络和蛋白表达模式的重建。这些改变的分子参与多种细胞过程，如蛋白水解、代谢、热休克反应、细胞骨架排列、氧化应激调节、翻译和信号转导。在TGF-β处理后，共有75个蛋白质点上调或下调超过两倍，其中42个通过质谱鉴定，包括主要的T. cruzi木瓜蛋白酶样半胱氨酸蛋白酶，在入侵中起重要作用，并参与寄生虫逃避宿主免疫系统的逃逸机制。在我们的研究中，我们观察到TGF-β的加入有利于上鞭毛体增殖，证实了2DE数据，其中先前描述的蛋白质参与这一过程被TGF-β正向刺激。
Transforming growth factor beta (TGF-β) plays a pivotal role in Chagas disease, not only in the development of chagasic cardiomyopathy, but also in many stages of the T. cruzi life cycle and survival in the host cell environment. The intracellular signaling pathways utilized by T. cruzi to regulate these mechanisms remain unknown. To identify parasite proteins involved in the TGF-β response, we utilized a combined approach of two-dimensional gel electrophoresis (2DE) analysis and mass spectrometry (MS) protein identification. Signaling via TGF-β is dependent on events of phosphorylation, which is one of the most relevant and ubiquitous post-translational modifications for the regulation of gene expression, and especially in trypanosomatids, since they lack several transcriptional control mechanisms. Here we show a kinetic view of T. cruzi epimastigotes (Y strain) incubated with TGF-β for 1, 5, 30 and 60 minutes, which promoted a remodeling of the parasite phosphorylation network and protein expression pattern. The altered molecules are involved in a variety of cellular processes, such as proteolysis, metabolism, heat shock response, cytoskeleton arrangement, oxidative stress regulation, translation and signal transduction. A total of 75 protein spots were up- or down-regulated more than twofold after TGF-β treatment, and from these, 42 were identified by mass spectrometry, including cruzipain–the major T. cruzi papain-like cysteine proteinase that plays an important role in invasion and participates in the escape mechanisms used by the parasite to evade the host immune system. In our study, we observed that TGF-β addition favored epimastigote proliferation, corroborating 2DE data in which proteins previously described to be involved in this process were positively stimulated by TGF-β.
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