FRET between CLAG3 and RhopH2 at distinct sites within infected cells. (A) Live-cell fluorescence microscopy of a C3R2-DKI schizont-infected cell before and after acceptor photobleaching (PB). Right panels show RGB color scaling of mCerulean fluorescence intensity; increased red labeling of individual rhoptries in the bottom panels indicates increased mCerulean intensity after mVenus
photobleaching. Color scale bar at right, 4 to 103 AU linear gradient. (B and C) Identical photobleaching experiments with LmC-R2mV negative-control parasite (RGB scale bar, 6 to 163 AU) and R3-C5V positive-control parasites (RGB scale bar, 0 to 298 AU). Images representative of 31 to 151 cells for each line. (E) Host membrane colocalization CLAG3-mCerulean and RhopH2-mVenus in a trophozoite-stage C3R2-DKI-infected cell. Colocalization at the host membrane is highlighted (boxed region and dashed line). (F) Corresponding line scan with yellow and cyan lines representing mVenus and mCerulean intensities. (H) Live-cell images of trophozoite-stage C3R2-DKI parasite before and after acceptor photobleaching to estimate FLIM-FRET. Right panels, RGB color scaling of mCerulean intensity (scale bar, 77 to 125 AU.
Ahmad M, Manzella-Lapeira J, Saggu G, Ito D, Brzostowski JA, Desai SA. Live-Cell FRET Reveals that Malaria Nutrient Channel Proteins CLAG3 and RhopH2 Remain Associated throughout Their Tortuous Trafficking. mBio. 2020 11(5):e01354-20.
Other associated proteins
|PF3D7_0929400||high molecular weight rhoptry protein 2|