Panigrahi Aswini K, Zíková Alena, Dalley Rachel A, Acestor Nathalie, Ogata Yuko, Anupama Atashi, Myler Peter J, Stuart Kenneth D
Seattle Biomedical Research Institute, Seattle, Washington 98109, USA.
Mol Cell Proteomics. 2008 Mar;7(3):534-45. doi: 10.1074/mcp.M700430-MCP200. Epub 2007 Dec 11.
African trypanosomes, early diverged eukaryotes and the agents of sleeping sickness, have several basic cellular processes that are remarkably divergent from those in their mammalian hosts. They have large mitochondria and switch between oxidative phosphorylation and glycolysis as the major pathways for energy generation during their life cycle. We report here the identification and characterization of several multiprotein mitochondrial complexes from procyclic form Trypanosoma brucei. These were identified and purified using a panel of monoclonal antibodies that were generated against a submitochondrial protein fraction and using tandem affinity purification (TAP) tag affinity chromatography and localized within the cells by immunofluorescence. Protein composition analyses by mass spectrometry revealed substantial divergence of oxidoreductase complex from that of other organisms and identified a novel complex that may have a function associated with nucleic acids. The relationship to divergent physiological processes in these pathogens is discussed.
非洲锥虫是早期分化的真核生物，也是昏睡病的病原体，它们具有几个基本的细胞过程，这些过程与它们的哺乳动物宿主的细胞过程显著不同。它们有大的线粒体，并且在生命周期中在氧化磷酸化和糖酵解之间切换，作为能量产生的主要途径。我们在此报告了从布氏锥虫前循环形式中鉴定和表征了几种多蛋白线粒体复合物。这些复合物是使用一组针对亚线粒体蛋白组分产生的单克隆抗体，并通过串联亲和纯化（TAP）标签亲和色谱法进行鉴定和纯化的，并通过免疫荧光在细胞内定位。通过质谱进行的蛋白质组成分析揭示了氧化还原酶复合物与其他生物体的显著差异，并鉴定出一种可能与核酸相关的新复合物。还讨论了这些病原体中不同生理过程之间的关系。
