PROTEOMIC ANALYSIS OF ZYGOTE AND OOKINETE STAGES OF THE AVIAN MALARIA PARASITE P

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 描绘的蛋白质组成的受精卵和动合子，早期发育阶段的疟原虫在蚊子中肠，是了解初始分子寄生虫-载体相互作用的基础。已发表的恶性疟原虫的蛋白质组不包括合子/动合子阶段的分析，伯氏疟原虫的蛋白质组也不包括合子阶段或分泌蛋白。制备了鸡胚丙酸杆菌受精卵、动合子和动合子分泌/释放的蛋白样品，并进行多维蛋白鉴定技术（MudPIT）。通过MS/MS鉴定鸡疟原虫受精卵、动合子和动合子分泌蛋白的肽，定位到鸡疟原虫全基因组序列的ORF（> 50个氨基酸），然后与恶性疟原虫中的同源ORF匹配。总共鉴定了966个编码orthopathy蛋白的恶性疟原虫ORF;发现这些预测的蛋白质中有40%以上是假设的。大多数具有预测的分泌信号肽或跨膜结构域的推定蛋白质是假设蛋白质。这项分析提供了一个迄今为止未知的蛋白质组的恶性疟原虫早期蚊子中肠阶段的更全面的看法。这些结果支持了对疟原虫-蚊子中肠相互作用的更强有力的研究，这对于开发阻断疟疾传播的新策略至关重要。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Delineation of the complement of proteins comprising the zygote and ookinete, the early developmental stages of Plasmodium within the mosquito midgut, is fundamental to understand initial molecular parasite-vector interactions. The published proteome of Plasmodium falciparum does not include analysis of the zygote/ookinete stages, nor does that of P. berghei include the zygote stage or secreted proteins. P. gallinaceum zygote, ookinete, and ookinete-secreted/released protein samples were prepared and subjected to Multidimensional protein identification technology (MudPIT). Peptides of P. gallinaceum zygote, ookinete, and ookinete-secreted proteins were identified by MS/MS, mapped to ORFs (> 50 amino acids) in the extent P. gallinaceum whole genome sequence, and then matched to homologous ORFs in P. falciparum. A total of 966 P. falciparum ORFs encoding orthologous proteins were identified; just over 40% of these predicted proteins were found to be hypothetical. A majority of putative proteins with predicted secretory signal peptides or transmembrane domains were hypothetical proteins. This analysis provides a more comprehensive view of the hitherto unknown proteome of the early mosquito midgut stages of P. falciparum. The results underpin more robust study of Plasmodium-mosquito midgut interactions, fundamental to the development of novel strategies of blocking malaria transmission.