Fig. 6

In vitro kinase assay of T. gondii component proteins with TgCaMKrk. a A schematic diagram of the TgGAP45 domain structure and its deletion mutant is shown. TgGAP45 consists of an N-terminal coiled-coil domain (shaded) and a C-terminal globular domain. The two phosphorylation sites, Ser¹⁶³ and Ser¹⁶⁷ [24], are indicated by arrows. The TgGAP45 mutant has a deletion in its coiled-coil domain (a.a. 27–136). b MBP-GAP45 mutant (predicted MW, 59 kDa; Lane 1), MBP-ACT1 (84 kDa; Lane 2), MBP-MLC1 (66 kDa; Lane 3), MBP-GAP50 (88 kDa; Lane 4), and MBP-LacZ (51 kDa; Lane 5) were immobilized on amylose resin beads and subjected to in vitro kinase assays with TgCaMKrk. The proteins were then washed three times with TNE buffer, separated on a denaturing gel, and stained with CBB. c An autoradiograph of the gel shown in A. Molecular masses (kDa) are indicated on the left