The mosquito stages of human-specific malaria parasites are difficult to study; however, in the avian-specific parasite Plasmodium gallinaceum, these stages are amenable to analysis. The transition between two such stages, zygote to ookinete, has been shown to be regulated by protein phosphorylation, but the specific proteins involved are not known. To identify proteins that are phosphorylated during this transition, a global phosphoproteomic analysis of the two stages will be conducted. The analysis will utilize a shotgun-proteomic approach termed MudPIT that separates and identifies proteins within complex mixtures by multidimensional liquid chromatography coupled to tandem mass spectrometry. Additionally, different techniques designed to enhance the detection of phosphorylation sites will be combined with MudPIT and compared. From this analysis, a chosen phosphoproteomic procedure will be used to identify phospho-regulated proteins that are involved in the transition between two obligate mosquito stages of the parasite, providing crucial data to the malaria research community that could result in the identification of candidate targets for blocking the transmission of malaria, a disease which affects the health of hundreds of millions of people worldwide.
