Expression analysis and co-localization of PFD1140w (A & B) with PfSBP-1 and PfEMP-1 at different stages of asexual parasite life cycle. Smears of methanol fixed Pf3D7-infected erythrocytes were stained with anti-PHIST antibodies (1:500), anti PfSBP-1 antibodies (1:1000) and anti PfEMP-1 antibodies (1:500), followed by incubation with Alexa flour-conjugated secondary antibodies (Alexa fluor 488 –green and Alexa fluor 546 – red). DIC: differential interference contrast image, DAPI: nuclear staining using DAPI (blue), PfEMP-1: mice anti-ATS antibody (red), PfSBP-1: anti-PfSBP-1 antibody (red), PFD1140w: rabbit anti-PFD1140w, antibodies (green), Merge: overlay of PHIST proteins with PfEMP-1 or PfSBP-1. Colocalization assays were performed with well-known MC marker, PfSBP-1 and surface expressed PfEMP-1 in different stages of parasite growth using their specific antisera. At trophozoite stage of parasite development, PFD1140w showed discrete punctuate pattern that appeared to co-localize with PfSBP-1 (A). In the schizont stage, significant overlapping of signals was observed in a punctuate organization within iRBC cytosol, suggestive of coexistence of PFD1140w and PfSBP-1 in MCs. Co-localization assays with PfEMP-1 displayed only partial overlap at the mature trophozoite stage of parasite development, while PfEMP-1 fluorescence seemed to surround PFD1140w fluorescence at schizont stages.
Kumar V, Kaur J, Singh AP, Singh V, Bisht A, Panda JJ, Mishra PC, Hora R.PHISTc protein family members localize to different sub-cellular organelles and bind Plasmodium falciparum major virulence factor 'PfEMP-1'. FEBS J. 2017 Nov 20.
Other associated proteins
|PF3D7_0424000||Plasmodium exported protein (PHISTc), unknown function|