Carbohydrate Recognition Specificity of Trans-sialidase Lectin Domain from Trypanosoma congolense
A: Schematic presentation of recombinant TconTS-LD fusion proteins expressed in bacteria. Fusion tags flanking TconTS-LD are: His: poly histidine tag, MBP: maltose binding protein tag, TEV: tobacco etch virus protease cleavage site, 3C: human rhinovirus 3C protease cleavage site, SNAP: SNAP-tag, Strep: Strep-tag. B: Homology model of TconTS2-LD comprising the α-helix calculated using TconTS2 amino acid sequence and crystal structure of Trypanosoma cruzi TS (PDB code: 3b69) as template employing the software Yasara. C: The molecular electrostatic surface of the homology model (B) was calculated using the ESPPME method of Yasara structure. Red colour indicates a positive potential, blue a negative and grey a neutral. A yellow ellipse indicates the groove encompassing the proposed binding site. D:SDS-PAGE of purified TconTS-LD proteins. After expression in E. coli Rosetta pLacI, 1–2 μg double affinity purified recombinant TconTS-LD, containing and lacking the α-helix, were loaded in each lane of an 10% SDS polyacrylamide gel as indicated. After electrophorese, the gel was stained using Coomassie Brilliant Blue.