Biophysical Feature, Crystallization and X-ray Crystallographic Studies of Toxascaris leonina Galectin

Abstract

Galectins are generally believed to be potential candidates for use in the development of novel antiinflammatory agents or as selective modulators of the immune response. In particular, galectin-9 exhibits some of the extracellular functions, including cell aggregation, adhesion, chemoattraction, activation, and apoptosis. Tl-galectin (Tl-gal, galectin-9 homologue gene) was isolated from an adult worm of the Toxascaris leonina. The full-length Tl-gal gene, which was incorporated into pET-28a, was overexpressed in E. coli and purified by nickel affinity and gel filtration chromatographies. The purified Tl-gal was crystallized using the hangingdrop vapor-diffusion method. The crystal belonged to the tetragonal space group <TEX>$P4_1$</TEX>, with unit-cell parameters of a = b = <TEX>$75.7\AA$</TEX> and c = <TEX>$248.4\AA$</TEX>. The crystals were obtained at <TEX>$20^{\circ}C$</TEX> and diffracted to a resolution of <TEX>$3.0\AA$</TEX>. The asymmetric unit contained four molecules of Tl-gal, which gave a crystal volume per protein mass (Vm) of <TEX>$2.8\AA^3Da^{-1}$</TEX> and a solvent content of 54.1%.