Queiroz Rayner M L, Charneau Sébastien, Bastos Izabela M D, Santana Jaime M, Sousa Marcelo V, Roepstorff Peter, Ricart Carlos A O
Department of Cell Biology, Institute of Biology, University of Brasilia , Brasília, Brazil.
J Proteome Res. 2014 Aug 1;13(8):3530-41. doi: 10.1021/pr401120y. Epub 2014 Jul 14.
Chagas' disease is a neglected infectious illness, caused by the protozoan Trypanosoma cruzi. It remains a challenging health issue in Latin America, where it is endemic, and so far there is no immunoprophylatic vaccine or satisfactory chemotherapic treatment for its chronic stage. The present work addressed the analysis of the plasma membrane (PM) subproteome from T. cruzi human-hosted life stages, trypomastigote and axenic amastigote, by two complementary PM protein enrichment techniques followed by identification using an LC-MS/MS approach. The results revealed an extensive repertoire of proteins in the PM subproteomes, including enzymes that might be suitable candidates for drug intervention. The comparison of the cell surface proteome among the life forms revealed some potentially stage-specific enzymes, although the majority was shared by both stages. Bioinformatic analysis showed that the vast majority of the identified proteins are membrane-derived and/or possess predicted transmembrane domains. They are mainly involved in host cell infection, protein adhesion, cell signaling, and the modulation of mammalian host immune response. Several virulence factors and proteins potentially capable of acting at a number of metabolic pathways of the host and also to regulate cell differentiation of the parasite itself were also found.
恰加斯病是一种被忽视的传染病,由原生动物克氏锥虫引起。在其为地方病的拉丁美洲,它仍然是一个具有挑战性的健康问题,而且到目前为止,对于其慢性阶段尚无免疫预防性疫苗或令人满意的化疗方法。本研究通过两种互补的质膜(PM)蛋白富集技术,随后采用液相色谱-串联质谱(LC-MS/MS)方法进行鉴定,对克氏锥虫在人体宿主中的生活阶段即锥鞭毛体和无共生体无鞭毛体的质膜亚蛋白质组进行了分析。结果揭示了质膜亚蛋白质组中广泛的蛋白质种类,包括可能是药物干预合适候选对象的酶。不同生活形态之间细胞表面蛋白质组的比较揭示了一些潜在的阶段特异性酶,尽管大多数酶在两个阶段都有。生物信息学分析表明,绝大多数已鉴定的蛋白质是膜衍生的和/或具有预测的跨膜结构域。它们主要参与宿主细胞感染、蛋白质黏附、细胞信号传导以及对哺乳动物宿主免疫反应的调节。还发现了几种毒力因子以及可能在宿主的多种代谢途径中起作用并调节寄生虫自身细胞分化的蛋白质。