Right: Transmission electron micrographs of Nluc-DH-APEX (parasites, expressing exported nanoluciferase) parasites showing membranous extensions induced by trapping protein cargo export. (A) Section through an Nluc-DH-APEX parasite treated with 10 nM WR99210 showing membranous extension indicated by arrows and enlarged in (B). Scale bars = 1 µm and 200 nm, respectively. (C) SIM immunofluorescence image of parasite labelled for Nluc and EXP2 showing that the large loops (arrows) seen in A,B are not exceptional. (D, E) Membranous extensions were not observed in Nluc-DH-APEX parasites not treated with WR99210. Scale bar = 1 µm. The infected erythrocytes were treated with the pore forming protein equinatoxin, to release and remove haemoglobin prior to fixation to prevent haemoglobin activating the peroxidase substrate. Scale bar = 1 µm. Left: HSP101 and HSP70-1 do not localise to the EXP2 PV loops formed in WR-treated Nluc-DH trophozoites. (A) Nluc-DH rings were treated with 10 nM WR and when trophozoites were fixed and probed with an EXP2 mouse monolconal and rabbit anti-HSP101 IgG. Widefield immunfluoroescence imaging reveals HSP101 localise to the PV (arrows) but not to the EXP2 PV loops. (B) Parasites similarly treated were also probed for the parasite cytoplasmic HSP70-1 protein which did not localise to the EXP2 labelled PV loops indicating they did not contain parasite cytoplasm. Scale bar = 5 µm.
Charnaud SC, Jonsdottir TK, Sanders PR, Bullen HE, Dickerman BK, Kouskousis B, Palmer CS, Pietrzak HM, Laumaea AE, Erazo AB, McHugh E, Tilley L, Crabb BS, Gilson PR. Spatial organisation of protein export in malaria parasite blood stages. Traffic. 2018 Apr 26. [Epub ahead of print] PMID: 29696751.
Other associated proteins
|PF3D7_1471100||exported protein 2|