Figure 6

(a) Recombinant PfTRAP interacts with tyrosine kinase proteins. HepG2 cell membrane extract were incubated with PfTRAP ECD. The complex was immunoprecipitated using PfTRAP antibodies, followed by western blot analysis using monoclonal antibodies against Lyn (i) (lane 2), Lck (ii) (lane 4), CrkL (iii) (lane 6). To check nonspecific interaction, HepG2 cell membrane extract alone was immunoprecipitated using PfTRAP antibodies (Lane 1, 3 and 5). (b) Recombinant PfTRAP is phosphorylated in the presence of CrkL, Lck and Lyn proteins. Lck (lane 2), Lyn (lane 4) and CrkL (lane 6) proteins were immunoprecipitated from HepG2 membrane extract and PfTRAP was incubated with the complex in the presence of γ33 ATP. PfTRAP was immunoprecipitated from the mixture and the phosphorylation was analyzed using autoradiography.