Recent Developments in Proteoglycan Purification and Analysis

Abstract

Proteoglycans are ubiquitous biomolecules in the body located in the extracellular matrix, on the cell surface and also within the cells. They contain at least one glycosaminoglycan (GAG) chain covalently attached to a core protein and may also present N-or O-linked glycans. The high structural diversity and distribution relate to the various biological functions of proteoglycans. In recent years, new members have enlarged the proteoglycan family and advances in molecular biology and glycobiology contributed to elucidate more of the biological functions of proteoglycans. In order to study the structure of a proteoglycan molecule and relate it to its function (or dysfunction), its isolation and purification from cell culture or tissue extracts is necessary. Next to the widely used anion exchange chromatographic methods, techniques based on lectin affinity chromatography have created new possibilities to increase the degree of purity. Introduction of electrospray ionization (ESI) and matrix-assisted laser desorption-ionization (MALDI) sources, together with tandem mass spectrometry (MS/MS or MS n ), mark a further important step towards the structural analysis of glycosaminoglycans. The aim of this review is to present the most recent advances in proteoglycan purification and analysis.

Glycosaminoglycans from Co-Products of «Scyliorhinus canicula»: Extraction and Purification in Reference to the European Pharmacopoeia Requirement

“…Analytical protocols for determining physicochemical characteristics and purity of the CS molecule are established. Several methods are commonly used for the extraction and segregation of GAGs [15, 30–32]. In this paper, all the methods of the monograph have been applied in addition to other methods that we suggested more accurate.…”

Proteoglycan sequence

“…Several detailed protocols for the preparation of GAGs and PGs for structure analysis have been recently published. 15,24,25 Slicing, grinding or pulverization of tissue samples is generally necessary for efficient PG extraction. After extraction, GuHCl and other salts present in the extraction buffer are generally removed by buffer exchange, prior to PG or GAG recovery from the extraction buffer using weak (W) anion-exchange (AX) or strong anion-exchange (SAX) chromatography on a column for large volume samples or a spin cartridge for small volume samples.…”

Structure elucidation of native N‐ and O‐linked glycans by tandem mass spectrometry (tutorial)

“…Other oligosaccharides such as those found in specific glycoconjugates, including glycolipids and peptidoglycans, have their own distinct fragmentation behavior. Furthermore, highly anionic oligosaccharides such as the glycosaminoglycans and the polysialic acids require specific chemical methods of pretreatment and specific methods for activation to obtain useful structural information (Zaia & Costello, 2003;Chi, Amster, & Linhardt, 2005;Didraga, Barroso, & Bischoff, 2006;Wolff et al, 2007).…”