Fig. 4.

Schistosoma mansoni venom allergen-like protein SmVAL6 interacts with both a fatty acid binding protein (Sm14) and a dynein light chain (DLC). (A) Sperm-Coating Protein/Tpx-1/Ag5/PR-1/Sc7 Protein/Tpx-1/Ag5/PR-1/Sc7 (SCP/TAPS)-containing SmVAL6 truncations T1 (143 aa; 32 kDa) and T2 (219 aa; 41 kDa) were generated from full-length SmVAL6 (Smp_124050) (a) and expressed in Saccharomyces] cerevisiae to identify adult worm interacting partners during yeast-2-hybrid (Y2H) screens (b). (B) The strength of SmVAL6/Sm14 DeltaE3 (Smp_095360.2) and SmVAL6/DLC (Smp_158660) interactions was quantified using both the pellet X-β-gal (PXG; black bars) (Mockli and Auerbach, 2004) and the Ortho-Nitrophenyl-β-galactoside (ONPG; grey bars) assays. Student’s t-tests were used to detect the significance of differential interactions quantified by the more sensitive PXG assay (SmVAL6T1/Sm14 DeltaE3 versus SmVAL6T2/Sm14 DeltaE3; SmVAL6T1/DLC versus SmVAL6T2/DLC). (C) Adult worms express alternatively spliced versions of Sm14: full-length Sm14 (Sm14 FL, Smp_095360.1) and the alternatively spliced Sm14 lacking exon 3 (Sm14 DeltaE3, Smp_095360.2). Gene structures are depicted where ‘E’ represents exons and arrows indicate PCR primers used to amplify both Sm14 isoforms (a). SCP/TAPS-containing SmVAL6 T1 interacts with both Sm14 isoforms as quantified by the PXG assay (b). ONPG and PXG controls included p53 + SV40 large T antigen (positive) and p53 + Lamin C (negative).