Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis*
Quantitative Proteomic and Phosphoproteomic Analysis of Trypanosoma cruzi Amastigogenesis*
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克氏锥虫无鞭毛发生的定量蛋白质组学和磷酸化蛋白质组学分析*
DOI：
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发表时间：
2014
影响因子：
7
通讯作者：
C. Ricart
中科院分区：
文献类型：
--
作者：
R. Queiroz;Sébastien Charneau;S. Mandacaru;V. Schwämmle;B. D. Lima;P. Roepstorff;C. Ricart
查加斯病是一种热带被忽视的疾病，在拉丁美洲流行，由原生动物克氏锥虫引起。该寄生虫有四个主要的生命阶段：外鞭毛体、后循环锥鞭毛体、血流锥鞭毛体和无鞭毛体。在酸性吞噬溶酶体中孵育一段时间后，感染性锥鞭毛体分化为复制性无鞭毛体，称为无鞭毛体发生。这种分化过程可以通过在酸性DMEM中孵育组织培养衍生的锥鞭毛体来体外模拟。在这里，我们使用这种成熟的分化方案进行全面的定量蛋白质组学和磷酸化蛋白质组学分析的T。cruzi无鞭毛发生对来自完全分化形式和两个生物学相关中间时间点的样品进行Lys-C/胰蛋白酶消化、iTRAQ标记和多重处理。随后，使用TiO 2基质富集磷酸肽。在LC-MS/MS分析之前，通过亲水相互作用液相色谱法分级分离非磷酸化肽。LC-MS/MS和生物信息学程序用于蛋白质和磷酸肽定量、鉴定和磷酸化位点分配。我们能够确定参与协调无鞭毛体发生的调节蛋白和途径。我们还观察到，一个显着比例的调节蛋白是膜蛋白。调节磷酸化事件协调的蛋白激酶和磷酸酶的一部分，在酸性介质中孵育诱导的信号级联反应也被证明。据我们所知，这项工作是迄今为止最全面的T. cruzi无鞭毛体发生，这些数据将作为未来研究的可靠基础，并可能成为新的潜在药物靶点。
Chagas disease is a tropical neglected disease endemic in Latin America caused by the protozoan Trypanosoma cruzi. The parasite has four major life stages: epimastigote, metacyclic trypomastigote, bloodstream trypomastigote, and amastigote. The differentiation from infective trypomastigotes into replicative amastigotes, called amastigogenesis, takes place in vivo inside mammalian host cells after a period of incubation in an acidic phagolysosome. This differentiation process can be mimicked in vitro by incubating tissue-culture-derived trypomastigotes in acidic DMEM. Here we used this well-established differentiation protocol to perform a comprehensive quantitative proteomic and phosphoproteomic analysis of T. cruzi amastigogenesis. Samples from fully differentiated forms and two biologically relevant intermediate time points were Lys-C/trypsin digested, iTRAQ-labeled, and multiplexed. Subsequently, phosphopeptides were enriched using a TiO2 matrix. Non-phosphorylated peptides were fractionated via hydrophilic interaction liquid chromatography prior to LC-MS/MS analysis. LC-MS/MS and bioinformatics procedures were used for protein and phosphopeptide quantitation, identification, and phosphorylation site assignment. We were able to identify regulated proteins and pathways involved in coordinating amastigogenesis. We also observed that a significant proportion of the regulated proteins were membrane proteins. Modulated phosphorylation events coordinated by protein kinases and phosphatases that are part of the signaling cascade induced by incubation in acidic medium were also evinced. To our knowledge, this work is the most comprehensive quantitative proteomics study of T. cruzi amastigogenesis, and these data will serve as a trustworthy basis for future studies, and possibly for new potential drug targets.
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布氏锥虫生命周期中酪氨酸磷酸化的独特模式。
影响因子：
1.5
作者：
Parsons,M;Valentine,M;Deans,J;Schieven,GL;Ledbetter,JA
通讯作者：
Ledbetter,JA
从溶酶体到细胞质：克氏锥虫的细胞内途径。
DOI：
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发表时间：
1994
期刊：
Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas
影响因子：
--
作者：
Andrews,NW
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Andrews,NW
影响因子：
--
作者：
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通讯作者：
Sailen Barik
DOI：
10.1016/j.bbapap.2005.08.018
发表时间：
2005-12-30
影响因子：
3.2
作者：
Naula, C;Parsons, M;Mottram, JC
通讯作者：
Mottram, JC
影响因子：
20.3
作者：
G. Pearson;Fred L Robinson;T. Gibson;Bing-e Xu;M. Karandikar;K. Berman;M. Cobb
通讯作者：
G. Pearson;Fred L Robinson;T. Gibson;Bing-e Xu;M. Karandikar;K. Berman;M. Cobb