Synthesis and conformational analysis of a pentasaccharide corresponding to the cell-wall polysaccharide of the Group A Streptococcus

Abstract

The synthesis and conformational analysis of a pentasaccharide corresponding to a fragment of the cell-wall polysaccharide (CWPS) of the bacteria Streptococcus Group A are described. The polysaccharide consists of alternating α-(1→2)- and α-(1→3)-linked l-rhamnopyranose (Rha p) residues with branching 2-acetamido-2-deoxy- d-glucopyranose (Glc pNAc) residues linked β-(1→3) to alternate rhamnose rings. The pentasaccharide is of interest as a possible terminal unit on the CWPS, for use in a vaccine. The syntheses employed a trichloroacetimidate glycosyl donor. Molecular dynamics ( md) calculations of the pentasaccharide with the force fields cvff and parm22, both in gas phase and with explicit water present, gave different predictions for the flexibility and preferred conformational space. Metropolis Monte Carlo ( mmc) calculations with the hsea force field were also performed. Experimental data were obtained from 1D transient NOE measurements. Complete build-up curves were compared to those obtained by full relaxation matrix calculations in order to derive a model of the conformation. Overall, the best fit between experimental and calculated data was obtained with mmc simulations using the hsea force field. Molecular dynamics and mmc simulations of a tetrasaccharide corresponding to the Group A-variant polysaccharide, which differs in structure from Group A in lacking the Glc pNAc residues, were also performed for purposes of comparison.