Human Blood Glycosaminoglycans: Isolation and Analysis

Abstract

Glycosaminoglycans (GAGs) are linear polysaccharides having disaccharide building blocks consisting of an amino sugar (N-acetylglucosamine, or N-acetylgalactosamine) and a uronic acid (glucuronic acid or iduronic acid) or galactose. Glycosaminoglycans have sulfated residues at various positions except for hyaluronan, and those sulfated residues regulate the biological functions of a wide variety of proteins, primarily through high-affinity interactions mediated by specific patterns/densities of sulfation and sugar sequences. Alteration of GAG structure is associated with a number of disease conditions and therefore the analyses of GAG structures and their sulfation patterns are important for the development of disease biomarkers and for treatment options. Extensive structural and quantitative analyses of GAGs from human blood are largely unexplored which may be due to the exhaustive isolation process because of the presence of too much interfering proteins and lipids such as serum albumin. Therefore we established a new GAG isolation method using the least amount (~200 μl) of human blood, consisting of a combination of proteolytic digestion and selective ethanol precipitation of GAGs, digestion of GAGs recovered on the filter cup by direct addition of GAG lyase reaction solution, and subsequent high-pressure liquid chromatography of unsaturated disaccharide products that enable to analyze GAG structures and contents. This isolation method offers an 80 % recovery of GAGs and can be applied to analyze a minute GAG content (≥1 nmol) from the least amount of biological fluids. Hence the method could be useful for the development of disease biomarkers.

Discrepancies in Composition and Biological Effects of Different Formulations of Chondroitin Sulfate

“…Chondroitin sulfate is one of five classes of glycosaminoglycans, the others being heparan sulfate, keratan sulfate, dermatan sulfate and hyaluronic acid [10]. Glycosaminoglycans are long, linear polysaccharides that contain a repeating disaccharide unit consisting of an amino sugar ( N -acetylglucosamine or N -acetylgalactosamine) and a uronic (glucuronic or iduronic) acid or galactose [11]. Various sulfated residues (except in hyaluronic acid) regulate their biological functions [11].…”

“…Glycosaminoglycans are long, linear polysaccharides that contain a repeating disaccharide unit consisting of an amino sugar ( N -acetylglucosamine or N -acetylgalactosamine) and a uronic (glucuronic or iduronic) acid or galactose [11]. Various sulfated residues (except in hyaluronic acid) regulate their biological functions [11]. Chondroitin sulfate [4] and hyaluronic acid [12] have been widely used in the treatment of osteoarthritis.…”

Chondroitin sulfate supplementation improves clinical outcomes in a murine model of necrotizing enterocolitis

“…Given that the sulfated residues of CS are believed to participate and regulate various biological functions (Anower & Kimata, 2015 ), as well as previous data from our lab suggesting that multiple other sulfur‐derived compounds exerted their beneficial properties through the eNOS pathway (Drucker, Jensen, Ferkowicz, & Markel, 2018 ; Hosfield et al, 2022 ; Jensen et al, 2017 ), we hypothesized that the effects of CS supplementation would also be mediated through this same pathway. The CS used for our experiments was bovine‐cartilage‐derived and mono‐sulfated, which was as expected from the literature (Martel‐Pelletier et al, 2015 ; Volpi, 2007 ).…”

“…Additionally, GAGs, and specifically CS, have been implicated in being a key component of the extracellular matrix which is lost in the intestines of neonates during NEC (Ade‐Ajayi et al, 1996 ). GAGs have sulfated residues that are believed to participate and regulate various biological functions (Anower & Kimata, 2015 ). CS can be a complex heterogenous polysaccharide with various sulfating patterns (du Souich et al, 2009 ; Martel‐Pelletier et al, 2015 ), but usually when extracted and processed from the cartilage of bovine, porcine, and galline sources, has single sulfate moieties (Martel‐Pelletier et al, 2015 ; Volpi, 2007 ).…”

A novel graphene oxide‐based fluorescence method for detection of urine glycosaminoglycans

“…1,3,6 Sulfated GAGs have an important role to play in the assessment of various diseases and serve as indicators and potential biomarkers. 7 The levels of GAGs in serum or urine are altered in several diseases such as cancer, Alzheimer's disease, and osteoarthritis. 8,9 Notably, sulfated GAGs are being determined for use in the diagnosis and treatment of mucopolysaccharidoses (MPSs) a type of lysosomal storage disorder.…”