Tracking Glideosome-associated protein 50 reveals the development and organization of the inner membrane complex of Plasmodium falciparum.
The most deadly of the human malaria parasites, Plasmodium falciparum, has different stages specialized for invasion of hepatocytes, erythrocytes, and the mosquito gut wall. In each case, host cell invasion is powered by an actin-myosin motor complex that is linked to an inner membrane complex (IMC) via a membrane anchor called the glideosome-associated protein 50 (PfGAP50). We generated P. falciparum transfectants expressing green fluorescent protein (GFP) chimeras of PfGAP50 (PfGAP50-GFP). Using immunoprecipitation and fluorescence photobleaching, we show that C-terminally tagged PfGAP50-GFP can form a complex with endogenous copies of the linker protein PfGAP45 and the myosin A tail domain-interacting protein (MTIP). Full-length PfGAP50-GFP is located in the endoplasmic reticulum in early-stage parasites and then redistributes to apical caps during the formation of daughter merozoites. In the final stage of schizogony, the PfGAP50-GFP profile extends further around the merozoite surface. Three-dimensional (3D) structured illumination microscopy reveals the early-stage IMC as a doubly punctured flat ellipsoid that separates to form claw-shaped apposed structures. A GFP fusion of PfGAP50 lacking the C-terminal membrane anchor is misdirected to the parasitophorous vacuole. Replacement of the acid phosphatase homology domain of PfGAP50 with GFP appears to allow correct trafficking of the chimera but confers a growth disadvantage.
La Trobe University, Biochemistry Department, Bundoora, Australia.
Jeffrey A
JA
Yeoman
Eric
E
Hanssen
Alexander G
AG
Maier
Nectarios
N
Klonis
Bohumil
B
Maco
Jake
J
Baum
Lynne
L
Turnbull
Cynthia B
CB
Whitchurch
Matthew W A
MW
Dixon
Leann
L
Tilley
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D052940:Merozoites / Q000502:physiology / Q000648:ultrastructure; D010963:Plasmodium falciparum / Q000166:cytology* / Q000378:metabolism* / Q000472:pathogenicity; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; C509918:glideosome-associated protein 45, Plasmodium falciparum; C509919:glideosome-associated protein 50, Plasmodium falciparum
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