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APEX2-based proximity proteomic analysis identifies candidate interactors for Plasmodium falciparum knob-associated
Sébastien Charneau1, Lucas Silva de Oliveira2,3, Zenon Zenonos4,5
1Laboratory of Biochemistry and Protein Chemistry, Department of Cell Biology, Institute of Biology, University of Brasília, Brasília, 70910-900, Brazil. charneau@unb.br.
Abstract:
The interaction of Plasmodium falciparum-infected red blood cells (iRBCs) with the vascular endothelium plays a crucial role in malaria pathology and disease. KAHRP is an exported P. falciparum protein involved in iRBC remodelling, which is essential for the formation of protrusions or "knobs" on the iRBC surface. These knobs and the proteins that are concentrated within them allow the parasites to escape the immune response and host spleen clearance by mediating cytoadherence of the iRBC to the endothelial wall, but this also slows down blood circulation, leading in some cases to severe cerebral and placental complications. In this work, we have applied genetic and biochemical tools to identify proteins that interact with P. falciparum KAHRP using enhanced ascorbate peroxidase 2 (APEX2) proximity-dependent biotinylation and label-free shotgun proteomics. A total of 30 potential KAHRP-interacting candidates were identified, based on the assigned fragmented biotinylated ions. Several identified proteins have been previously reported to be part of the Maurer's clefts and knobs, where KAHRP resides. This study may contribute to a broader understanding of P. falciparum protein trafficking and knob architecture and shows for the first time the feasibility of using APEX2-proximity labelling in iRBCs.
Insights
Researchers identified 30 proteins interacting with KAHRP in Plasmodium falciparum-infected red blood cells (iRBCs). This study advances understanding of malaria parasite protein trafficking and knob formation, crucial for disease pathology.
Area of Science:
- Malariology
- Cell Biology
- Parasitology
Background:
- Plasmodium falciparum-infected red blood cells (iRBCs) interact with the endothelium, contributing to malaria pathology.
- Knobs on iRBC surfaces, mediated by proteins like KAHRP, are vital for cytoadherence and immune evasion but can cause severe complications.
Purpose of the Study:
- To identify proteins interacting with KAHRP, a key protein in iRBC remodelling and knob formation.
- To explore the feasibility of APEX2-proximity labelling in iRBCs for protein interaction studies.
Main Methods:
- Utilized enhanced ascorbate peroxidase 2 (APEX2) proximity-dependent biotinylation.
- Employed label-free shotgun proteomics to analyze protein interactions.
- Applied genetic and biochemical tools to investigate KAHRP interactions.
Main Results:
- Identified 30 potential KAHRP-interacting protein candidates.
- Confirmed known proteins associated with Maurer's clefts and knobs among the identified candidates.
- Demonstrated the successful application of APEX2-proximity labelling in iRBCs.
Conclusions:
- The study provides a list of potential KAHRP interactors, aiding the understanding of knob architecture.
- This research contributes to understanding Plasmodium falciparum protein trafficking mechanisms.
- APEX2-proximity labelling is a viable method for studying protein interactions in iRBCs.
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