Distinct Roles of Plasmodium Rhomboid 1 in Parasite Development and Malaria Pathogenesis
Figure 5
PbTRAP is not a substrate for PbROM1.
(A) PbTRAP is cleaved by a serine protease. Total cell lysate from 3×104 salivary gland sporozoites was loaded in each lane of a 4–25% denaturing SDS-PAGE. Western blot was performed using an anti-PbTRAP-repeat rabbit polyclonal antibody. The TRAP fragment recognized by the anti-repeat antibody but not by antibody against the cytoplasmic tail is indicated with an asterisk. Ctrl, control; E, empty lane. (B) PbTRAP processing in PbROM1(−) sporozoites. Experiments with 3×104 sporozoites/lane were conducted as described for experiments in panel A with wild-type (WT) and mutant parasites. (C) Gliding motility of PbROM1(−) sporozoites. PbROM1(−) salivary gland sporozoites were placed in a 2-well chamber slide coated with BSA and incubated at 37°C for 30 min. After fixation with paraformaldehyde CSP trails were detected with an anti-CSP antibody (3D11) and a rhodamine-conjugated anti-mouse secondary antibody. *, sporozoite at the leading end of the trail.