Group B IMC associated proteins reveal distinct dynamics. A. The GFP-tagged alveolins PF10_0039, PFE1285w and MAL13P1.228 emerge as large ring-like structures (T1-2) that towards the end of schizogony show the same subpellicular distributions as their TMD counterparts (T3).Kono M, Herrmann S, Loughran NB, Cabrera A, Engelberg K, Lehmann C, Sinha D, Prinz B, Ruch U, Heussler V, Spielman T, Parkinson J, Gilberger TW. Evolution and Architecture of the Inner Membrane Complex in Asexual and Sexual Stages of the Malaria Parasite. Mol Biol Evol. 2012 29(9):2113-32.

Upper panel: Co-localization of the group A protein PFD1110w (anti-PFD1110w, red) with the group B alveolin PFE1285w (PFE1285w-GFP, green) emphasizes the differential nature of the two structures in the nascent IMC and their spatial relation during the ongoing schizogony (T1-3). Nuclei stained with DAPI. Scale bar 2 μm.Lower panel: A double transgenic parasite line was generated expressing a different combination of proteins, MAL13P1.130-GFP (group A) and PF10_0039-mCherry. The dynamic distribution of these two members confirmed the spatial distinction but intimate proximity of the group A and group B defined IMC sub-compartments during the early phase of IMC biogenesis.Kono M, Herrmann S, Loughran NB, Cabrera A, Engelberg K, Lehmann C, Sinha D, Prinz B, Ruch U, Heussler V, Spielman T, Parkinson J, Gilberger TW. Evolution and Architecture of the Inner Membrane Complex in Asexual and Sexual Stages of the Malaria Parasite. Mol Biol Evol. 2012 29(9):2113-32

Subcellular distribution of proteins predicted to be involved in invasion. All proteins were localized in schizonts (s) and free merozoites (m) using GFP-fusion proteins and grouped according to their predominant GFP localization: surface (green), inner membrane complex (turquoise),Hu G, Cabrera A, Kono M, Mok S, Chaal BK, Haase S, Engelberg K, Cheemadan S, Spielmann T, Preiser PR, Gilberger TW, Bozdech Z. Transcriptional profiling of growth perturbations of the human malaria parasite Plasmodium falciparum. Nat Biotechnol. 2010 28(1):91-8.

IFA was performed on PfCDPK1-3HA-DD parasites using anti-HA antibody to detect CDPK1 and anti-IMC1g antisera. Scale bar is 2 μm. Data strongly suggest that PfCDPK1 is a major kinase that phosphorylates IMC1g in the parasite. Moreover, PfCDPK1 exhibited localization in close proximity with IMC1g in the parasite, which was reflected by overlapping fluorescence in IFA.Kumar S, Kumar M, Ekka R, Dvorin JD, Paul AS, Madugundu AK, Gilberger T, Gowda H, Duraisingh MT, Keshava Pr asad TS, Sharma P. PfCDPK1 mediated signaling in erythrocytic stages of Plasmodium falciparum. Nat Commun. 2017;8(1):63.

Colocalization of BTP1 with ALV5 and GAPM2 during inner membrane complex (IMC) biogenesis. (A) Colocalization of BTP1–GFP with ALV5–mCherry (red) in unfixed parasites. The initial ring structure marked by BTP1–GFP precedes the appearance of Alveolin 5 (ALV5) (T1). The emerging Alveolin-5-defined ring is associated but distinct from that marked by BTP1 (T2). During IMC expansion, BTP1 remains rim associated (T3) and, at the end of daughter cell formation, is concentrated at the basal end with ALV5 equally distributed within the IMC (T4). (C) Colocalization of BTP1–GFP with GAPM2 (red) in fixed parasites. Both proteins appeared at about the same time, marking the onset of IMC biogenesis (T1). GAPM2 stains small ring–shaped formations that are encircled by BTP1–GFP (T2). With elongation and propagation of the IMC from the apical towards the basal end of the forming daughter cell, GAPM2 is equally distributed within the forming IMC, but BTP1 is only present at the rim of the nascent IMC (T3) and, at the end of schizogony, is concentrated at the basal pole of the daughter cell (T4). Nuclei were stained with DAPI (blue). Enlargement of selected areas are marked with a white square and referred to as ‘zoom’ (a fourfold magnification). The bottom row shows a schematic representation of the spatio-temporal distribution of BTP1 and GAPM2. Kono M, Heincke D, Wilcke L, Wong TW, Bruns C, Herrmann S, Spielmann T, Gilberger TW. Pellicle formation in the malaria parasite. J Cell Sci. 2016 129(4):673-80.