Apicomplexan parasites such as Plasmodium falciparum and Toxoplasma gondii cause malaria and toxoplasmosis, respectively, which afflict millions worldwide. In T. gondii, a lysine (K) methyltransferase (KMT) localises to the apical complex in tachyzoites and regulates parasite egress. This enzyme, termed apical complex KMT (TgAKMT), is an SET-domain-containing KMT with orthologues in other apicomplexan species, including PfSET7 in P. falciparum. Both enzymes localise to the parasite’s apical complex and play key roles in regulating motility. Owing to their distinct structural features and essential cellular roles, AKMTs represent promising targets for anti-parasitic drug development.
PfSET7 was crystallised, and its structure was determined to ~2.1 Å resolution. It adopts a tertiary fold highly similar to TgAKMT and forms homodimers in a concentration-dependent manner. Uniquely, PfSET7 contains a beta-hairpin within the SET domain, which is conserved only in orthologues belonging to the Hematozoa subclass, such as Babesia and Theileria spp. Deletion of the hairpin drastically reduces enzymatic activity, whereas substitution with the homologous beta-hairpin from Theileria parvum preserves function. PfSET7 also contains four conserved motifs within its intrinsically disordered N-terminal region, found exclusively in Plasmodium spp., which may enable liquid-liquid phase separation, as suggested by our preliminary studies. The cellular functions of both of these distinct PfSET7 features remain to be elucidated.
All AKMT orthologues share a conserved, unique pocket adjacent to the substrate lysine channel, presenting an attractive site for inhibitor binding. A high-throughput screen of ~70,000 compounds identified six hits that specifically inhibit PfSET7 activity. These initial hits are currently being optimised to generate more potent and selective inhibitors.
Together, our studies will deepen understanding of this unique KMT family and apicomplexan biology. In parallel, the drug discovery aspect of this work holds promise for delivering novel therapeutic strategies against malaria, toxoplasmosis, and related diseases.