A comparative proteomic approach to identify and validate African trypanosome proteins at the host-parasite interface

寄生虫病困扰着世界上的许多人？S人口，以及地球上大多数人处于一种或几种这样的病原体的危险之中。这些寄生虫引起的疾病范围从疟疾到肠道、皮肤和器官感染。在非洲，锥虫寄生虫造成了许多死亡和发病--感染是在受感染的采采蝇进食血餐时造成的。如果不治疗，感染者将通过一般的健康不佳症状进展到多器官功能障碍、昏迷和死亡。相关的锥虫也会在亚洲、南美洲、中东和南欧引起疾病。由于非洲锥虫定期更换皮毛的特殊系统，宿主免疫系统无法控制寄生虫，这会导致有效的感染，如果不治疗，必然会死亡。由于外衣不同，疫苗不太可能接种，而且使用的药物超过50年，往往有毒，由于出现耐药性而失败。迫切需要新的研究来确定潜在的新治疗选择和诊断方法。寄生虫独一无二的一个方面是表面的一个区域，在那里形成毛皮的所有分子在形成时都会出现，称为鞭毛袋。由于表面是宿主和寄生虫之间的边界，了解这种表面是如何维持的具有重大的科学意义和实用价值，有助于发现锥虫可能对药物治疗敏感的新方式。我们希望找出鞭毛袋中存在哪些蛋白质，期望对存在的分子的了解将为了解鞭毛袋如何发挥作用提供洞察力。到目前为止，定义鞭毛口袋的成功有限，我们提出了一种方法，需要相当大的计算机能力和广泛的分析来解决这个问题。最重要的是，在确定候选鞭毛口袋蛋白之后，我们将寻找哪些预期的蛋白已经确定，并将结合计算机预测，使用水母绿色荧光蛋白或GFP定位一系列未知蛋白。这将使我们能够评估有多少蛋白质真正是鞭毛袋的一部分，从而开始了解这个细胞器如何有助于锥虫致病的能力。

Parasitic diseases afflict many of the world?s population, and the majority of people on our planet at at risk from one or several such agents. The diseases these parasites cause range from malaria to gut, skin and organ infections. In Africa, the trypanosome parasite is responsible for much death and morbidity - infection is brought about when an infected tsetse fly takes a blood meal. Without treatment the infected person will progress through general ill health symptoms to multiple organ malfunction, coma and death. Related trypanosomes also cause disease in Asia, South America, the Middle East and Southern Europe.Due to a remarkable system where the African trypanosome changes its coat at regular intervals, the host immune system is unable to control the parasite, which results in efficient infection and invariably death if untreated. Vaccines are unlikely due to the varying coat, and the drugs in use at over 50 years old, frequently toxic and failing due to emergence of resistance. Urgent new research is needed to identify potential new therapeutic options and diagnostics. One aspect of the parasite that is unique is a region of the surface where all molecules that form the coat emerge when they are made, called the flagellar pocket. As the surface is the boundary between host and parasite, understanding how this is maintained is of major scientific interest as well as of practical utility in uncovering new ways in which the trypanosome may be vulnerable to drug treatments. We wish to find out what proteins are present in the flagellar pocket, with the expectation that an understanding of the molecules that are present will provide insights into how the pocket performs its job. There has so far been limited success with defining the flagellar pocket, and we propose a method that requires considerable computer power and wide-raging analysis as a solution to the problem. Most importantly, following on from the identification of candidate flagellar pocket proteins, we will look to see which expected proteins have been identified, and also, in combination with computer predictions, will localize a range of unknown proteins using the jellyfish green fluorescent protein, or GFP. This will allow us to assess how many of the proteins really are parts of the flagellar pocket, and hence begin to understand how this organelle contributes to the ability of the trypanosome to cause disease.