title: On the association between amyloid fibrils and glycosaminoglycans; possible interactive role of Ca2+ and amyloid P-component.
abstract: We have previously reported the specific association of glycosaminoglycans (GAG) and proteoglycans (PG) with amyloid fibrils and characterized the polysaccharides directly extracted from amyloid-laden tissues. In the present study we further elucidate the association between purified amyloid fibrils and GAG/PG with special reference to those GAG/PG associated with amyloid P-component (AP) and the interactive role of Ca2+ ions. Amyloid fibrils were isolated from human hepatic AA amyloid employing water extraction with and without preceding removal of AP, an extrafibrillar protein component of all amyloids, using sodium citrate. GAG/PG co-isolated with the amyloid extracts, with and without AP, were isolated and characterized. Agarose-affinity chromatography of extracts containing AP was performed, and the GAG associated with this extrafibrillary protein were characterized as well. Several different GAG/PG populations were demonstrated in the various extracts. The abolition of calcium-dependent binding markedly influenced the amount of GAG/PG recovered in the fibril extracts, as well as the total amount of amyloid material obtained. Thus, it seems that calcium plays an important role in the association between the fibrils and the sugar moieties, and that a significant fraction of the GAG found in amyloid exhibits a Ca(2+)-dependent fibril-GAG interaction. No significant difference in the proportion between galactosaminoglycans and glucosamines was, however, disclosed when the two extraction protocols were compared, suggesting that no particular GAG species has a higher affinity for the fibrils themselves. Both dermatan/chondroitin sulphate and heparan sulphate identified in the present study exhibited a Ca(2+)-dependent interaction with AP, supporting previous findings. However, the amyloid-associated galactosaminoglycans found, especially the large PG appearing in small amounts, seemed to have a higher affinity for the extrafibrillar AP than the other GAG.
authors.affiliation: Department of Rheumatology, University of Tromsø, Norway.
authors.forename: T
authors.initials: T
authors.lastname: Stenstad
authors.forename: J H
authors.initials: JH
authors.lastname: Magnus
authors.forename: K
authors.initials: K
authors.lastname: Syse
authors.forename: G
authors.initials: G
authors.lastname: Husby
mesh_terms: D000682:Amyloid / Q000032:analysis* / Q000302:isolation & purification; D002118:Calcium / Q000502:physiology*; D002648:Child; D006025:Glycosaminoglycans / Q000032:analysis* / Q000302:isolation & purification; D006801:Humans; D008297:Male; D011509:Proteoglycans / Q000032:analysis / Q000302:isolation & purification; D000683:Serum Amyloid P-Component / Q000502:physiology*
chemical_list: D000682:Amyloid; D006025:Glycosaminoglycans; D011509:Proteoglycans; D000683:Serum Amyloid P-Component; D002118:Calcium