rPfNPT1 is phosphorylated by rPfCDPK5 in vitro. In vitro kinase reactions were performed with [32P]ATP using recombinant versions of PfNPT1 and PfCDPK5. BSA was used as a negative control. (B) PfNPT1 localizes to the parasite plasma membrane. Schizont stage parasites + Shld1 were treated with E64, fixed, and probed with anti-HA (PfCDPK5), anti-V5 (PfNPT1), anti-PfGAP45, and/or anti-PfMSP1 antibodies. Nuclei were stained with DAPI (4’,6-diamidino-2-phenylindole). Scale bar, 1 mm. PfCDPK5-3HA localization both shows diffusion within the parasite cytoplasm and displays increased signal near the parasite plasma membrane. Costaining with anti-V5 and antihemagglutinin (anti-HA) antibodies revealed a partial overlap PfCDPK5 and PfNPT1, particularly in areas near the parasite plasma membrane (top row). PfGAP45 is a protein associated with the inner membrane complex which underlies the plasma membrane. Costaining with anti-PfGAP45 and anti-V5 showed partial overlap (middle row). However, with antibodies to the parasite plasma membrane protein PfMSP1, we detected strong colocalization of PfNPT1 and PfMSP1 (bottom row), demonstrating that PfNTP1 localizes to the parasite plasma membrane.
Blomqvist K, Helmel M, Wang C, Absalon S, Labunska T, Rudlaff RM, Adapa S, Jiang R, Steen H, Dvorin JD. Influence of Plasmodium falciparum Calcium-Dependent Protein Kinase 5 (PfCDPK5) on the Late Schizont Stage Phosphoproteome. mSphere. 2020 Jan 8;5(1). pii: e00921-19
Other associated proteins
|PF3D7_0930300||merozoite surface protein 1|
|PF3D7_1222700||glideosome-associated protein 45|
|PF3D7_1337800||calcium-dependent protein kinase 5, CDPK5|