PfPNPLA2 localizes to the mitochondria during different parasites stages. (A) A putative patatin-like phospholipase, PfPNPLA2 (PF3D7_1358000) contains a FaD-like domain (green) and a patatin-like phospholipase domain (yellow) along with the GXSXG-motif and catalytic aspartate characteristic of this family of proteins. (B) Downregulation of PfPNPLA2-HA levels. Schizont cultures of the PfPNPLA2-HA-KD parasite line were treated with 2.5 mM GlcN for 72 h to determine knockdown of PfPNPLA2. Lysates were immunoblotted with rat anti-HA antibody to detect PfPNPLA2-HA (~240 kDa); immunoblotting with rabbit anti-Pf39 antisera (39 kDa) was used as loading control (bottom panel). Quantification of PfPNPLA2-HA levels following knockdown. PfPNPLA2-HA levels were evaluated between the GlcN-treated and untreated PfPNPLA2-HA-KD parasites in three independent Western blots based on the band intensity as estimated with ImageJ software (right panel). Pf39 levels were used for normalization (set to 100%). (C) Using a mouse anti-PfPNPLA2 antibody (green) for immunolabelling, PfPNPLA2 is detectable in perinuclear regions in schizonts and gametocytes of stages II-V. Counter labeling of the asexual blood stages was done using rabbit anti-PfMSP1 antisera and gametocytes with rabbit anti-Pfs230 antisera (red). The nuclei were labeled with Hoechst 33342 nuclear stain (blue). Bar; 5 µm. (D) PfPNPLA2-HA, as detected by rat anti-HA antibody (green) does not colocalize to PfPNPLA3 (red), which was immunolabelled using a mouse anti-PfPNPLA3 antibody, and (E) PfPNPLA4-HA (green), detected by rat anti-HA antibody, does not localize with PfPNPLA2, immunolabelled with a mouse anti-PfPNPLA2 antibody (red). (F) PfPNPLA2 has a Mitotracker™ (red) co-localization during asexual stages using PfPNPLA2-HA-glmS parasites line using an anti-HA antibody (green). (G) Live microscopy using Mitotracker (red), revealed that the loss of PfPNPLA2 leads to degradations of the mitochondrion morphology in trophozoite stage of the parasite.

Shunmugam S, Quansah N, Flammersfeld A, Islam MM, Sassmannshausen J, Bennink S, Yamaryo-Botté Y, Pradel G, Botté CY. The patatin-like phospholipase PfPNPLA2 is involved in the mitochondrial degradation of phosphatidylglycerol during Plasmodium falciparum blood stage development. Front Cell Infect Microbiol. 2023 13:997245. PMID: 38089812