Isolation and separation of proteoglycans

Abstract

Proteoglycans contain a polypeptide core and an oligosaccharide chain composed of aminohexoses and uronic acid. The glycan chain is attached to the polypeptide in a bond to serine hydroxyl. The glycan chains may contain up to 200 disaccharide units and the proteoglycan molecular mass ranges from a few thousands to millions. Their physiological functions are related to barriers limiting diffusion across the membranes, articular lubrification, blood coagulation and cellular adhesion. The tissue proteoglycans can be extracted with 4 M guanidine hydrochloride and purified with chromatographic techniques. The soluble proteoglycans can be precipitated with cetylpyridinium chloride, purified by chromatography or by dialysis. All proteoglycan species are amenable to electrophoresis on polyacrylamide gels, and after blotting on polyvinylidene fluoride membranes, they can be stained for glycans. Proteoglycan analyses have shown their value in clinical mucopolysaccharidosis diagnostics, in occupational toxicology and in coagulation studies. Experimental applications include cell adhesion studies in tumor biology, regeneration in neurosciences or maturation of skin and kidneys.