Synthesis and solution structure of an S-glycosylated cyclic hexapeptide. Evidence for conformational change induced by glycosylation

Synthesis and conformational analysis of the S-glycosylated cyclic hexapeptide cyclo(-D-Pro1-Phe2-Cys3(tetra-O-acetyl-beta-D-galactopyranosyl)-++ +Trp4-Lys(Z)5- Phe6-) I was carried out to examine the influence of a saccharide residue in position i of a standard beta-turn on the formation of reverse turns and on the biological activity. Synthesis was carried out in the liquid phase employing a galactosylated cysteine building block. The cyclization reagents DPPA/NaHCO3 avoided high dilution conditions. Spectroscopic data were extracted from homo- and heteronuclear 2D-NMR techniques (TOCSY, NOESY, HMQC, HMQC-TOCSY, HMBCS-270). For structural refinement restrained molecular dynamics (MD) simulations in vacuo and with explicit DMSO as solvent were performed. Finally, simulations in DMSO without experimental restraints provided insight in stability and dynamics of the structural model. A comparison of the S-glycosylated Cys3 peptide with the analogous Thr3 peptide exhibits a similar overall conformation of the hexapeptide [beta II' D-Pro-Phe and another beta-turn about Trp4-Lys5(Z)]. However, the latter shows a distinct dynamic flip beta I, beta II in the glycopeptide, whereas the Thr-analogue only populates beta I. This influence is attributed to a beta I stabilizing effect of a hydrogen bridge of Thr-O gamma in position i to the NH of the amino acid in position i + 2, which is lacking in the glycosylated compound.