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. 2015 Jan 12;112(4):1179–1184. doi: 10.1073/pnas.1415466112

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Fig. 3. — PfRH5/PfRipr/CyRPA complex is GPI-anchored to the merozoite surface. (A) 3D reconstruction of IFA z-stacks during merozoite invasion, coimmunostained with MSP-1 detected PfRH5, PfRipr, and CyRPA at the apical end of the merozoite surface. In the 3D images, the γ settings were altered for visual representation only. (B) Triton X-114 separates PfRipr and CyRPA (detergent fraction) from PfRH5 (aqueous fraction). (C) CyRPA has the ω-site (SSS); weak hydrophobic tail as observed in known GPI-anchored parasite proteins, which PfRipr lacks. Arrow denotes the predicted GPI:protein-transamidase cleavage site, where GPI is attached. (D) Immunoprecipitation of tritium-radiolabeled CyRPA from lysates of schizont-stage parasites grown in the presence of tritium labeled GPI anchor precursors d-[6-³H(N)]glucosamine hydrochloride and d-[2-³H]mannose confirmed the presence of a GPI anchor. (E) Immunoblot analysis of anti-CyRPA coimmunoprecipitated proteins (same radiolabeled sample) confirmed the presence of PfRH5/PfRipr/CyRPA.