Discovery of Platyhelminth-Specific α/β-Integrin Families and Evidence for Their Role in Reproduction in Schistosoma mansoni
Figure 5
Interaction studies confirmed binding of Smβ-Int1 to the schistosome cellular kinases SmTK3, SmTK6, and SmTK4. A:
For binding studies in the YTH system, yeast cells (strain AH109) were co-transformed with the prey plasmid Smβ-Int1-C-term pACT2 together with the baits SmTK3-SH4SH3 pBridge, SmTK3-SH3 pBridge, SmTK6-SH4SH3 pBridge, SmTK6-SH3 pBridge, and SmTK4-SH2SH2 pBridge. Yeast clones were selected on Trp−/Leu−/His− media (T−/L−/H−) for interactions between bait and prey proteins, and β-galactosidase colony lift filter assays were performed for detection of LacZ expression. B: Comparative β-galactosidase liquid assays were performed with the yeast clones from A to determine the relative binding strengths. As control, untransformed yeast cells (AH109) were used (control). The statistical evaluation of seven independent measurements of β-Gal activity (n = 7) is shown (error bars are indicated). C: Co-Immunoprecipitation of HA-Smβ-Int1 and V5-SmTK4 expressed in Xenopus oocytes. Anti-HA antibodies immunoprecipitated Smβ-Int1 together with SmTK4 upon co-expression in oocytes. Inversely, anti-V5 antibodies immunoprecipitated SmTK4 with Smβ-Int1, when they were expressed together in oocytes.