CK2 Secreted by Leishmania braziliensis Mediates Macrophage Association Invasion: A Comparative Study between Virulent and Avirulent Promastigotes

Figure 3

Partial purification of the sCK2 in the virulent sample of L. braziliensis. (a) Identification of kinase activity. All of the obtained fractions were assayed for CK2 activity as described in Section 2 [32]. Inset. Western blotting of the purified secreted CK2 of the virulent L. braziliensis sample. Fraction 45 was submitted to 12% SDS-PAGE (Lane 1), transferred to a PVDF membrane (Lane 2), incubated with an anti-CK2 antibody, and developed as described in Section 2. (b) Phosphorylation of the supernatant proteins of the virulent Leishmania braziliensis sample by the CK2 enzyme secreted by this parasite. After the supernatant was obtained in the absence (control) or presence (experimental) of dephosphorylated casein, the assay was performed as described in Section 2. (1) Proteins stained with Coomassie Brilliant Blue R250. (2) Phosphorylated proteins observed by exposure to a phosphorimager plate (a: control supernatant, b: experimental supernatant, c: control supernatant + TBB, d: experimental supernatant + TBB, e: control supernatant + BIS, f: control supernatant + heparin, and g: experimental supernatant + heparin).

(a)

(b)