MODELING THE CARBOHYDRATE-BINDING SPECIFICITY OF PIG EDEMA TOXIN

The wild-type binding pentamer of Shiga-like toxin IIe (SLT-IIe) binds both the globotriaosylceramide (Gb(3)) and globotetraosylceramide (Gb (4)) cell surface glycolipids, whereas the double mutant GT3 (Q65E/K67 Q) exhibits a marked preference for Gb(3) [Tyrrell, G. J., et al. (199 2) Proc. Natl. Acad. Sci. U.S.A. 89, 524-528]. We modeled three unique sites (sites 1-3) for binding of the carbohydrate moiety of Gbs to GT 3 and SLT-IIe, on the basis of the three sites observed for the SLT-I pentamer [Ling, H., et al. (1998) Biochemistry 37, 1777-1788]. Examina tion of the three sites in light of various mutation and binding data strongly suggested that one of the binding sites plays a role in the c hange of specificity observed for the GT3 mutant. We applied several m odeling techniques, and developed a model for binding of the carbohydr ate moiety of Gb(4) to this site of the SLT-IIe binding pentamer. This model is consistent with a wide variety of mutation and binding data and clearly shows the importance of the terminal GalNac residue of Gb( 4), as well as that of the two mutated residues of GT3, to the intermo lecular interaction.