Hugo Belda

Functional analysis of kinases secreted into the host cell by the human malaria parasite Plasmodium falciparum

Hugo Belda - 3rd year PhD


The major focus will be the phenotypic characterisation of conditional knockout mutant parasites for 4 exported FIKK kinases with a view to gleaning insights into the function of this enigmatic group of putative enzymes. The knockouts will be subjected to a range of assays examining the surface structure, biophysical properties and cytoadherence characteristics of the parasitised cells, as well as using phosphoproteomics to examine effects of gene disruption on phosphorylation of parasite and host cell components.
In addition a recombinant version of the kinase domain of these FIKK kinases will be generated in order to identify potential inhibitors. The most potent of the inhibitors identified from the PKIS library show antiparasite activity in vitro, and efforts will be made to determine whether these are on-target only or due to off-target effects, as well as examining their potency against Plasmodium knowlesi (which lacks most of the FIKK kinases). Recombinant expression of additional members of the selected FIKK subgroup will be attempted, and artificial substrates will be sought. It was recommended that attempts could be usefully made (in collaboration) to obtain structural information on the recombinant kinases and on the inhibitors.