Independent Analysis of the Flagellum Surface and Matrix Proteomes Provides Insight into Flagellum Signaling in Mammalian-infectious Trypanosoma brucei
Independent Analysis of the Flagellum Surface and Matrix Proteomes Provides Insight into Flagellum Signaling in Mammalian-infectious Trypanosoma brucei
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DOI：
10.1074/mcp.m111.010538
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发表时间：
2011-10-01
影响因子：
7
通讯作者：
Hill, Kent L.
中科院分区：
文献类型：
--
作者：
Oberholzer, Michael;Langousis, Gerasimos;Hill, Kent L.
非洲锥虫的鞭毛是一种重要的多功能细胞器，在运动，细胞形态发生和宿主-寄生虫相互作用中发挥作用。以前的研究锥虫鞭毛已受到限制，无法纯化鞭毛，而不首先去除鞭毛膜。这种限制在研究鞭毛信号传导的背景下特别相关，因为信号传导需要鞭毛膜中的表面暴露蛋白和鞭毛基质中的可溶性信号传导蛋白。在这里，我们采用了遗传和机械方法相结合，以纯化完整的鞭毛从非洲锥虫，布氏锥虫，在其寄生虫感染阶段。我们结合鞭毛纯化与亲和纯化的表面暴露的蛋白质进行独立的蛋白质组学分析的鞭毛表面和基质馏分。鉴定的蛋白质涵盖广泛的分子功能，包括许多预测在信号传导中起作用的蛋白质。免疫荧光和RNA干扰研究表明鞭毛定位和功能的蛋白质鉴定和鞭毛附着和运动机制提供了深入的了解。鞭毛表面蛋白质组包括许多T.布鲁氏菌特异性蛋白质，并富含在寄生虫生命周期的疟原虫感染阶段上调的蛋白质。综合结果表明，鞭毛表面提出了一个多样化的和动态的主机-寄生虫接口，非常适合主机-寄生虫信号。Molecular & Cellular Proteomics 10：10.1074/mcp. M111.010538，1-14，2011.
The flagellum of African trypanosomes is an essential and multifunctional organelle that functions in motility, cell morphogenesis, and host-parasite interaction. Previous studies of the trypanosome flagellum have been limited by the inability to purify flagella without first removing the flagellar membrane. This limitation is particularly relevant in the context of studying flagellum signaling, as signaling requires surface-exposed proteins in the flagellar membrane and soluble signaling proteins in the flagellar matrix. Here we employ a combination of genetic and mechanical approaches to purify intact flagella from the African trypanosome, Trypanosoma brucei, in its mammalian-infectious stage. We combined flagellum purification with affinity-purification of surface-exposed proteins to conduct independent proteomic analyses of the flagellum surface and matrix fractions. The proteins identified encompass a broad range of molecular functionalities, including many predicted to function in signaling. Immunofluorescence and RNA interference studies demonstrate flagellum localization and function for proteins identified and provide insight into mechanisms of flagellum attachment and motility. The flagellum surface proteome includes many T. brucei-specific proteins and is enriched for proteins up-regulated in the mammalian-infectious stage of the parasite life-cycle. The combined results indicate that the flagellum surface presents a diverse and dynamic host-parasite interface that is well-suited for host-parasite signaling. Molecular & Cellular Proteomics 10: 10.1074/mcp.M111.010538, 1-14, 2011.