Plasmodium falciparum infection induces dynamic changes in the erythrocyte phospho-proteome

Abstract

The phosphorylation status of red blood cell proteins is strongly altered during the infection by the malaria parasite Plasmodium falciparum. We identify the key phosphorylation events that occur in the erythrocyte membrane and cytoskeleton during infection, by a comparative analysis of global phospho-proteome screens between infected (obtained at schizont stage) and uninfected RBCs. The meta-analysis of reported mass spectrometry studies revealed a novel compendium of 495 phosphorylation sites in 182 human proteins with regulatory roles in red cell morphology and stability, with about 25% of these sites specific to infected cells. A phosphorylation motif analysis detected 7 unique motifs that were largely mapped to kinase consensus sequences of casein kinase II and of protein kinase A/protein kinase C. This analysis highlighted prominent roles for PKA/PKC involving 78 phosphorylation sites. We then compared the phosphorylation status of PKA (PKC) specific sites in adducin, dematin, Band 3 and GLUT-1 in uninfected RBC stimulated or not by cAMP to their phosphorylation status in iRBC. We showed cAMP-induced phosphorylation of adducin S59 by immunoblotting and we were able to demonstrate parasite-induced phosphorylation for adducin S726, Band 3 and GLUT-1, corroborating the protein phosphorylation status in our erythrocyte phosphorylation site compendium.

Analysis of erythrocyte signalling pathways during Plasmodium falciparum infection identifies targets for host-directed antimalarial intervention

“…In addition, phosphorylation of Ser695 may also be involved in the localization of PKCθ to the plasma membrane 27 . These data are consistent with previous reports that host cell PKC activity is detectable during schizont development (30-40 h post invasion) with limited activity before 25 h 23,24 , and considerably refine these previous findings, notably with respect to dynamic phosphorylation changes on specific residues of the various PKC isoforms during parasite development.…”

Signalome-wide assessment of erythrocyte response toPlasmodiumreveals novel targets for host-directed antimalarial intervention

“…Additionally, phosphorylation of Ser695 may also be involved in the localisation of PKCθ to the plasma membrane 27 . These data are consistent with previous reports that host cell PKC activity is detectable during schizont development (30-40 hrs post-invasion) with limited activity before 25 hrs 23,24 , and considerably refine these previous findings, notably with respect to dynamic phosphorylation changes on specific residues of the various PKC isoforms during parasite development.…”

“…Some of the most notable changes implicated the PKC isoforms δ and θ. Several PKC isoforms have previously been reported to be activated during P. falciparum infection of erythrocytes, but the phosphorylation status of these kinases was uncharacterised 23,24 . Several phosphorylation sites of PKCδ/θ were significantly phosphorylated in trophozoite- and schizont-infected cells (Fig.…”

Interaction of Plasmodium falciparum casein kinase 1 with components of host cell protein trafficking machinery

“…None of these phosphopeptides map to the Vps9 domain or the Ras‐GAP domain (Figure 1c), suggesting that they probably do not directly regulate GTP turnover (although allosteric effects cannot be excluded), but likely function in regulating other essential functions of GAPVD1, such as adaptor protein interactions. Five of the eight phosphosites (T391, S903, S904, S972, and S999) have been previously described in GAPVD1 from human and mouse cells, 33,34 including S903 (corresponding to PfCK1 residue S902) in uRBCs 35 . The other three phosphosites (Y461, S915, and S1104) are hitherto undescribed in GAPVD1 proteins (Table 1).…”