Extended Data Fig. 1: PfPTRAMP and PfCSS interact with the PfRh5 complex and play an essential role for growth of P. falciparum.
From: PCRCR complex is essential for invasion of human erythrocytes by Plasmodium falciparum
a-f. Volcano plots illustrating the log2 fold change (log2FC) of immunoprecipitated HA-tagged CyRPA, PfRipr, or PfRh5 versus 3D7 before and after cross-linking with DSP versus significance of the change (-log10 P value). Differential protein expression analysis was performed using Limma which involves a moderated t-test. Benjamini and Hochberg’s method was used to adjust the p-values for multiple testing. Proteins were deemed differentially regulated in the log2 fold change in protein expression was 1-fold and exhibited an adjusted p-value ≤ 0.05. Proteins that were significantly immunoprecipitated with PfCyRPA-HA, PfRipr-HA and PfRh5-HA were analysed further and this included PfPTRAMP, PfCSS and Apical Membrane Protein 1 (AMA1). In this study we concentrated on PfPTRAMP and PfCSS. We are testing a potential link of AMA1 with the function of the PCRCR complex and this will be published elsewhere. An additional protein that was significantly found in all three immunoprecipitation experiments was heat shock protein 70 (PF3D7_0917900) but this was not considered further because it is a highly expressed chaperone protein and frequently immuno-precipitates in experiments such as these. Proteins that immunoprecipitated significantly in less than the three conditions were analysed with respect to subcellular location, timing of expression and potential role in merozoite invasion and because they did not match these criteria were not analysed further. a. Immuno-precipitated HA-tagged CyRPA. b. Immunoprecipitated HA-tagged PfRipr. c. Immuno-precipitated HA-tagged PfRh5. d. Immunoprecipitated HA-tagged CyRPA after cross-linking proteins with DSP. e. Immuno-precipitated HA-tagged PfRipr after cross-linking proteins with DSP. f. Immuno-precipitated HA-tagged PfRh5 after cross-linking proteins with DSP. g. Parasitemia of P. falciparum parasites with rapamycin inducible knockdown of PfPTRAMP. h. Parasitemia of P. falciparum parasites with rapamycin inducible knockdown of PfCSS. i. Parasitemia of P. falciparum parasites with rapamycin inducible knockdown of PfRh5.
