Fig. 7: NBPXa acts prior to moving junction formation and can be synergistically targeted with DBP. | Nature Communications

Fig. 7: NBPXa acts prior to moving junction formation and can be synergistically targeted with DBP.

From: Sequential roles for red blood cell binding proteins enable phased commitment to invasion for malaria parasites

Fig. 7

a IFAs of Pk merozoites stalled mid-invasion with cyto D. Top panels depict AMA1-mNG + NBPXa-HA. Bottom panels depict DBPα-mNG + AMA1-HA. Overlays stained with wheatgerm agglutinin (Magenta). Scale bars = 2 μm. b Processed NBPXa is detected by western blot in culture supernatants (S) when egressing merozoites are allowed to re-invade fresh RBCs and (c) when invasion is blocked with anti-DARC (α-D). Cont = control IgG. For all blots, NBPXa was detected with rat anti-HA (targeting C-term) and/or rabbit anti-NBPXa raised against the NBPXa N-terminus (amino acids 151-467). Anti-PfHSP70 used as loading control. P = pellet fraction. Green region in schematic shows transmembrane ___domain, * indicates putative ROM4 cleavage site. d Clustal alignment of several predicted RBL/DBP transmembrane domains. All contain a conserved alanine residue (red box), followed by putative helix destabilising motifs underlined in blue, which may serve as ROM4 recognition sites. All sequences apart from NBPXa from Baker et al.33 & O’Donnell et al.31. The experimentally determined PfEBA-175 cleavage position is indicated with black arrow. (O’Donnell et al.31). Parasite growth inhibition activity of anti-NBPXa (e) and anti-DB10 (f) was assessed individually and in combination (e) using fixed DB10 concentration with increasing anti-NBPXa concentrations Results calculated from two independent experiments. Error bars indicate ±SEM. Source data for (b), (c), (e) and (f) are provided as a Source Data file.

Back to article page