Interactome and reverse genetics studies of regulatory subunits of Plasmodium falciparum casein kinase

Malaria is the most devastating parasitic infection killing each year between 1 and 3 million of people. Furthermore this decease has a huge socio-economic impact in affected countries. The emergence of resistance against currently used drug makes the development of a new treatment an urgent task. Considering the success of kinase inhibitors in treatment of various cancers and considering the divergence between the parasites and human kinome, proteins kinases have become an attractive potential target. In this project we have used mass spectrometry analysis to study the Interactome of three P.flaciparum kinases: PfCK2beta1; PfCK2beta2 and PfGSK3. The genetically modified lines that we have used possess haemaglutinin tag. We have used this haemaglutinin tag to immunoprecipitate our proteins and their potentials interactors. Samples were then separated on acrylamide gel and analysed at the proteomics core facility at EPFL. Finally we have analysed the potential interactors and study the pathway in which they are involved. Using these methods we have found that the three PfCK2 subunits were immunoprecipitate together. These were already deemed known results since it has already been shown that these subunits interact together. But since there is, to the best of our knowledge, no example of this kind of study in P.falciparum, this project tends to demonstrate that mass spectrometry can be used to study Interactome of the parasites proteins. For our three proteins of interest we have identified some potential interactors but our results are too preliminary to conclude that those proteins are real interactors. A better understanding of the parasite life cycle and its metabolic pathway is critical in the development of a treatment against Malaria