Expression and localization of Kae1cyto interaction partners and Sua5. Representative IFA images from RFA-tagged parasites are shown. Localization for all three proteins is consistent with cytoplasmic localization, and Bud32 (top row), Cgi121 (middle row), and Sua5 (bottom row) show diffuse staining throughout the parasite. Nuclei were visualized with DAPI-DNA stain. Mallari JP, Oksman A, Vaupel B, Goldberg DE. Kinase-Associated Endopeptidase 1 (Kae1) Participates in an Atypical ribosome-Aassociated complex in the apicoplast of Plasmodium falciparum. J Biol Chem. 2014 Sep 9 [Epub ahead of print]

Expression and localization of Kae1api and Kae1cyto. Representative images of trophozoites are shown. Kae1api (top row) co-localizes with an apicoplast marker (acyl carrier protein, ACP), while Kae1cyto (second row) displays a diffuse signal throughout the parasite. Mitochondria and nuclei werevisualized with Mitotracker and DAPI-DNA stain, respectively. Lower row: Immuno-EM of Kae1api demonstrates localization to the apicoplast, bound by multiple membranes. Bar scale = 0.5 μm. Kae1api) displayed an elongated, punctate signal overlapping precisely with that of the apicoplasttargetedacyl carrier protein (ACP) in trophozoites and early schizonts . Both signals were detected in a compartment adjacent to, but distinct from the mitochondrion, consistent with apicoplast localization. Immuno-EM primarily detected Kae1api within structures bound by 3-4 membranes, further supporting that it is an apicoplast-targeted protein. In contrast, IFA detected a diffuse signal for Kae1cyto) throughout the parasite, suggesting a cytoplasmic location.Mallari JP, Oksman A, Vaupel B, Goldberg DE. Kinase-Associated Endopeptidase 1 (Kae1) Participates in an Atypical ribosome-Aassociated complex in the apicoplast of Plasmodium falciparum. J Biol Chem. 2014 Sep 9 [Epubahead of print]