Structurally defined synthetic heparin oligosaccharides reveal unique signatures for nanopore structural analysis of GAGs

Abstract

International audienceEfforts to fully understand the structure-to-function relationships of glycosaminoglycans (GAGs) have beenhampered by the complexity and polydispersity of GAGs and, more importantly, by the need for sensitiveanalytical techniques to discern their subtle structural motifs. Conventional analytical methods often fail toprovide information on the sparse motifs of GAGs. Concurrently, nanopore-based analysis has shown great potentialin GAGs structural analysis. Still, until now, it has been limited by the difficulty of relating ionic currentchanges to specific motifs. To address this, the present study allowed for obtaining reference signals byemploying carefully designed synthetic sulfated tetra- to dodecasaccharides to allow precise sequence controland purity. Reference signals were obtained for specific motifs, paving the way for accurate motif recognition.The outcomes of the study demonstrated unequivocal recognition of specific GAG structures by monitoring theevolution of nanopore signals in response to the translocation of synthetic heparin (HP)-like oligosaccharideswith controlled structures, allowing extensive structural discrimination. The findings highlight the nanopore'scapability to detect subtle structural features, such as differences in sulfation patterns and epimerization ofuronic acid. These results open the way to establishing detailed heparin (HP)-specific reference signals and tonanopore fingerprinting and sequencing of GAGs