title: Identification of a role for the PfEMP1 semi-conserved head structure in protein trafficking to the surface of Plasmodium falciparum infected red blood cells.
abstract: Transport of Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1) variants to the red blood cell (RBC) surface enables malarial parasite evasion of host immunity by modifying the antigenic and adhesive properties of infected RBCs. In this study, we applied the Bxb1 integrase system to integrate transgenes encoding truncated PfEMP1-GFP fusions into cytoadherent A4 parasites and characterize their surface transport requirements. Our studies revealed that the semi-conserved head structure of PfEMP1 proteins, in combination with the predicted transmembrane region and cytoplasmic tail, encodes sufficient information for RBC surface display. In contrast, miniPfEMP1 proteins with truncated head structures were exported to the RBC cytoplasm but were not detected at the RBC surface by flow cytometry or immuno-electron microscopy. We demonstrated the absence of a mechanistic barrier to having native and miniPfEMP1 proteins displayed simultaneously at the RBC surface. However, surface-exposed miniPfEMP1 proteins did not convey cytoadherence properties to their host cells, implicating potential steric considerations in host-receptor interactions or the need for multiple domains to mediate cell binding. This study establishes a new system to investigate PfEMP1 transport and demonstrates that the PfEMP1 semi-conserved head structure is under selection for protein transport, in addition to its known roles in adhesion.
authors.affiliation: Seattle Biomedical Research Institute, Seattle, WA, 98109, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Melcher
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Muhle
authors.forename: Philipp P
authors.initials: PP
authors.lastname: Henrich
authors.forename: Susan M
authors.initials: SM
authors.lastname: Kraemer
authors.forename: Marion
authors.initials: M
authors.lastname: Avril
authors.forename: Ines
authors.initials: I
authors.lastname: Vigan-Womas
authors.forename: Odile
authors.initials: O
authors.lastname: Mercereau-Puijalon
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Smith
authors.forename: David A
authors.initials: DA
authors.lastname: Fidock
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D017124:Conserved Sequence; D004912:Erythrocytes / Q000469:parasitology*; D005434:Flow Cytometry; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D010963:Plasmodium falciparum / Q000235:genetics / Q000378:metabolism*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C085731:erythrocyte membrane protein 1, Plasmodium falciparum; D049452:Green Fluorescent Proteins