Localization of wild type and mutant PfDJ1 in parasites. Wild type P. falciparum 3D7 parasites (anti-DJ1) were assessed for expression of native DJ1 by IFA using anti-PfDJ1 antibodies. Live recombinant parasites were evaluated for expression of GFP/DJ1WT (DJ1WT) or mutants (C106A, S151A and P154A) using a fluorescence microscope. The images show the presence of native DJ1 or GFP/DJ1 specific signal (DJ1), staining of the nucleus (Nuc), parasite and the infected erythrocyte boundaries (Bright), and all three images overlapped (Merged). Intense diffuse signal in all the parasites indicates expression of DJ1 proteins. Fluorescence microscopy of live parasites confirmed expression of GFP/DJ1 fusion proteins (Fig. 8), which was similar to that of native DJ1 in wild type parasites and the previously reported localization of native DJ1.
Nair DN, Prasad R, Singhal N, Bhattacharjee M, Sudhakar R, Singh P, Thanumalayan S, Kiran U, Sharma Y, Sijwali PS. A conserved human DJ1-subfamily motif (DJSM) is critical for anti-oxidative and deglycase activities of Plasmodium falciparum DJ1. Mol Biochem Parasitol. 2018 222:70-80. .