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LC-MS/MS based proteomic analysis of in vivo cap-binding protein and its interacting-partners in Plasmodium falciparum using Impact II

ABSTRACT: The emergence and spread of Plasmodium falciparum human malaria parasites resistant to antimalarial drugs, including artemisinin, a first line antimalarial drug, is threatening malaria treatment and prompting fears of a resurgence of the disease. New drugs with novel mode of actions are thus urgently required. Several compounds with antimalarial activity are known to target protein translation, although few of these targets have been validated. Translation initiation in eukaryotes is known to require eukaryotic translation initiation factor 4F (eIF4F) complex, which binds to the 5′-cap structure on mature mRNA and recruits other proteins for translation of mRNA. The putative components of the eIF4F complex in P. falciparum have been identified in the genome including PfeIF4E, a 5′-cap-binding protein; PfeIF4A, a helicase protein for unwinding mRNA, and PfeIF4G, a PfeIF4E/PfeIF4A scaffold protein, which could constitute a novel antimalarial target. However, it is not known if these proteins constitute a P. falciparum eIF4F complex in vivo, nor what other proteins interact with the mRNA 5′-cap to control translation initiation in this species. Here, we investigated P. falciparum proteins that interact with the mRNA 5′-cap. Native protein extract from P. falciparum parasites was applied to m7GTP agarose beads and specific binding proteins eluted using m7GTP. LC-MS/MS based proteomic analysis of the m7GTP-eluted proteins demonstrated the presence of PfeIF4E, which was not found in control experiments with non-methylated GTP beads, verifying the native cap-binding function of PfeIF4E. PfeIF4A, PfeIF4G, and a putative polyadenylate-binding protein-interacting protein were present among m7GTP-eluted proteins but in low abundances. Interestingly, proteomics data clearly demonstrated P. falciparum enolase (Pfeno) in the m7GTP-eluted proteins.

ORGANISM(S): Plasmodium falciparum 3D7  

SUBMITTER: Aiyada Aroonsri 

PROVIDER: PXD025553 | JPOST Repository | Fri Apr 22 00:00:00 BST 2022


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