Evaluation of Intermolecular Association of Glycosaminoglycan Oligosaccharides Using Nanoelectrospray Ionization Mass Spectrometry

Abstract

This study examines the non-covalent interactions between glycosaminoglycan (GAG) oligosaccharides using nanoelectrospray ionization mass spectrometry (nanoESI-MS). It is the first time that interactions between oligosaccharides have been observed using MS. The importance of interactions between GAGs has recently attracted much interest because they are related to biological functions. For instance, hyaluronic acid (HA) is known to associate with chondroitin sulfates (CSs), although the details of the interaction remain unclear. In general, non-covalent interactions between glycans are too weak to detect by general means. In this work, we applied nanoESI-MS with high sensitivity, which is widely used to observe non-covalent interactions, to investigate the interaction between HA and CSs. HA and CS oligosaccharides are used to discuss the results in a simplified manner. Our approach is aimed at interpreting the behavior of GAG polysaccharides from the information obtained using the oligosaccharides. HA and CS tetrasaccharides were demonstrated to associate to form heterodimer ions that were easily detected using nanoESI-MS. We also determined the stoichiometry of the interaction and calculated the Kd values of the interactions between HA and CS tetrasaccharides. How these structures affect the strength and stability of the non-covalent complexes is discussed. Further study of the interactions between HA and CS oligosaccharides will clarify the biological meaning of the coexistence of HA and CS in body fluids and tissues.