Composition of proteoglycan fragments from hyaline cartilage produced by granulocytes in a model of frustrated phagocytosis.
An in vitro model of frustrated phagocytosis was developed in which granulocytes interact with well-defined slices of hyaline cartilage. The composition of the purified proteoglycan fragments released from the cartilage slices by N-formyl-methionyl-leucyl-phenylalanine-stimulated granulocytes was studied after 30, 60 and 90 min incubation time. It was shown that the proteoglycan fragments do not change their composition during incubation. The only change observed during incubation was an increase in the quantity of the fragments. The protein content of the proteoglycan fragments is 7.0-8.6%, corresponding to a peptide chain of 24-28 amino acids, and the relative molecular mass of the total fragment is Mr = 37,600-39,200. On average, each proteoglycan fragment contains two chondroitin sulphate chains (Mr = 22,000-22,400), every fourth fragment contains a keratan sulphate chain (Mr = 7000-7200) and every seventh to eighth contains an O-glycosidic oligosaccharide, whereas no N-glycosidic oligosaccharide could be detected. The results of the disaccharide analysis show that the galactosaminoglycan chains contain 76.2-83.6% chondroitin 4-sulphate, 12.9-19.4% chondroitin 6-sulphate, 3.5-3.8% chondroitin and no dermatan sulphate. Since composition and relative molecular mass of the chondroitin sulphate and keratan sulphate chains from the proteoglycan fragments resemble those of native proteoglycans, the conclusion may be drawn that the degeneration of the proteoglycans occurs by proteases that attack preferably the chondroitin sulphate-rich region of the core protein. This is the first inflammation model of joint destruction, which demonstrates the elution of soluble specific proteoglycan degradation products of defined size.
Liszt
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Institut für Klinische Chemie und Pathobiochemie, RWTH Aachen.
Schnittker-Schulze
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Stuhlsatz
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Greiling
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D000596:Amino Acids / Q000032:analysis; D002356:Cartilage; D002809:Chondroitin Sulfates / Q000737:chemistry* / Q000302:isolation & purification; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004187:Disaccharides / Q000302:isolation & purification; D006098:Granulocytes / Q000502:physiology*; D006595:Hexosamines / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D008954:Models, Biological; D008970:Molecular Weight; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009300:Nasal Septum; D009504:Neutrophils / Q000187:drug effects / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry* / Q000302:isolation & purification; D010587:Phagocytosis / Q000187:drug effects; D011509:Proteoglycans / Q000378:metabolism*
D000596:Amino Acids; D004187:Disaccharides; D006595:Hexosamines; D010446:Peptide Fragments; D011509:Proteoglycans; D009240:N-Formylmethionine Leucyl-Phenylalanine; D002809:Chondroitin Sulfates