Left: Confocal microscopy localizes PfAnkDHHC (palmitoyl transferase) to Golgi. 3D7 schizonts were labeled with (A) rat anti-PfAnkDHHC, and (B) rabbit anti-ERD2, a known Golgi marker. (C) Merged image of the two stains shows extensive co-localization. (D) Corresponding DIC image. All images 630x.Right: Proposed model of topology of PfAnkDHHC in the Golgi membrane. The DHHC domain is exposed to the cytosol to serve as a palmitoyl transferase to cytosoloic proteins or transmembrane proteins with cysteines.Seydel KB, Gaur D, Aravind L, Subramanian G, Miller LH. Plasmodium falciparum: characterization of a late asexual stage golgi protein containing both ankyrin and DHHC domains. Exp Parasitol. 2005 110:389-93. Copyright Elsevier 2010.

PfDHHC1 is an IMC localized palmitoyl acyltransferase. E. PfDHHC1-GFP was localized in unfixed late stage parasites showing characteristic IMC dynamics during schizogony (T1-T3). Nuclei stained with DAPI (blue). Enlargement of selected areas are marked with a white square and referred as zoom. Scale bar, 1 μm. F. Co-localization of PfDHHC1-GFP and PfISP3mCherry. PfISP3mCherry was episomally co-expressed and shows identical spatial distribution compared with PfDHHC1-GFP. G. PfDHHC1-GFP distribution within the nascent IMC during gametocytog-enesis (stage IV and V). These symmetric sutures of the IMC are expanding with ongoing growth of the IMC vesicles and maturation of the gametocytes (stage III and IV). Scale bars, 1 μm.Wetzel J, Herrmann S, Swapna LS, Prusty D, Peter AT, Kono M, Saini S, Nellimarla S, Wong TW, Wilcke L, Ramsay O, Cabrera A, Biller L, Heincke D, Mossman K, Spielmann T, Ungermann C, Parkinson J, Gilberger TW. The role of palmitoylation for protein recruitment to the inner membrane complex of the malaria parasite. J Biol Chem. 2014 Nov 25. PMID:

Over-expression and localization of PfDHHC1, PfDHHC2, PfDHHC3 and PFDHHC9 in late stage parasites. A. Expression of PfDHHC1-GFP. A2. Localization of PfDHHC1-GFP in unfixed parasites showing characteristic IMC dynamics during schizogony: Commencing as cramp-like structures (T1), transforming to small ring-shaped formations (T2) that towards the end of schizogony expand and are then equally distributed underneath the plasma membrane (T3). Nuclei are stained with DAPI (blue). Enlargement of selected areas are marked with a white square and referred to as zoom. Scale bar, 1 μm. A3. Co-localization with the IMC marker GAPM2 (anti-GAPM2, red) in fixed cells confirmed PfDHHC1-GFP IMC localization shown here in an early stage of TMC biogenesis (T1). B. Expression of PfDHHC2-GFP. B2. Live microscopy of PfDHHC2-GFP revealed a circular structure around the nucleus of the nascent merozoites reminiscent of the ER. Scale bar, 1 μm. Wetzel J, Herrmann S, Swapna LS, Prusty D, Peter AT, Kono M, Saini S, Nellimarla S, Wong TW, Wilcke L, Ramsay O, Cabrera A, Biller L, Heincke D, Mossman K, Spielmann T, Ungermann C, Parkinson J, Gilberger TW. The role of palmitoylation for protein recruitment to the inner membrane complex of the malaria parasite. J Biol Chem. 2014 Nov 25. PMID:

Confocal microscopy demonstrates the location of PfAnkDHHC posterior to apical organelles. (A) 3D7 merozoites were labeled with rat-antiPfAnkDHHC and Alexa-488 conjugated anti-rat IgG (green) and with anti-RAP-1 and Alexa-594 conjugated anti-mouse IgG (red). Nuclei were stained with DAPI and appear blue. (B) DIC image. (C) 3D7 merozoites in schizonts labeled with anti-PfAnkDHHC (green) and a mouse antisera recognizing subtilisin-2 (red). Merozoites attached to the pigment containing residual bodies have their apical end oriented opposite to the residual body. (D) DIC image corresponding to Fig. 2C. All images 630x.Seydel KB, Gaur D, Aravind L, Subramanian G, Miller LH. Plasmodium falciparum: characterization of a late asexual stage golgi protein containing both ankyrin and DHHC domains. Exp Parasitol. 2005 110:389-93. Copyright Elsevier 2010.