Figure 1: EhCaBP1 binds EhC2PK in vitro and in vivo.
From: A C2 domain protein kinase initiates phagocytosis in the protozoan parasite Entamoeba histolytica
(a) Schematic presentation of domain organization of EhC2PK. The protein has a molecular mass of 50 kDa and two domains, C2 (1–91 amino-acid residues) and protein kinase (150–408 amino-acid residues). The active site of the kinase was predicted to reside within 269–281 amino-acid residues (arrow). (b) Immunoprecipitation of EhCaBP1 from whole-cell lysate of E. histolytica using CNBr-conjugated anti-C2PK antibody. EhC2PK was also immunoprecipitated from the lysate using CNBr-conjugated anti-C2PK antibody as a control. The total input lysate was also probed for the presence of EhC2PK, EhCaBP1 and EhCaBP2 by their respective antibodies. (c) Co-precipitation of EhCaBP1 or its calcium-binding defective mutant EhCaBP1ΔEF with wild-type GST-EhC2PK and GST-K179A mutant. In vitro GST-tagged kinase domain alone (GST-KD) was not able to precipitate EhCaBP1. GST alone did not bind to EhCaBP1. Glutathione Sepharose beads were used for precipitation experiments. All molecular markers are in kDa.
