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

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 描述蚊子中肠内疟原虫早期发育阶段的受精卵和动子组成的蛋白质组成的补体，是了解最初的分子寄生虫-媒介相互作用的基础。已发表的恶性疟原虫蛋白质组不包括合子/运动阶段的分析，伯氏疟原虫的蛋白质组也不包括合子阶段或分泌蛋白。制备鸡疫霉受精卵、动细胞和动细胞分泌/释放的蛋白质样品，并进行多维蛋白质鉴定技术(MudPIT)。用MS/MS鉴定鸡疟原虫受精卵、动子和动子分泌蛋白的多肽，定位到鸡疟原虫全基因组序列中的ORFs(&gt；50个氨基酸)，然后与恶性疟原虫的同源ORFs进行比对。总共鉴定出966个恶性疟原虫ORF编码同源蛋白；这些预测蛋白中略高于40%被发现是假设的。大多数具有预测的分泌信号肽或跨膜区的推定蛋白质是假想蛋白质。这一分析提供了迄今为止未知的恶性疟原虫早期蚊子中肠阶段蛋白质组的更全面的看法。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，这对开发阻止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.