Characteristics of Proteoglycans Containing Chondroitin and Dermatan Sulfates in the Mineral and Matrix Phases of Rabbit Bone

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Rabbit long bone proteoglycans (PGs) were extracted with guanidine-HCI (GdnCDI), EDTA and then with GdnCI to demonstrate their localization. The PGs in the EDTA extract (E-extract) and second GdnCl extract (G2-extract) were concentrated on an anion-exchange column, and their characteristics were determined by Western blot analysis after electrophoresis. Chondroitin 4-sulfate (C4S), chondroitin 6-sulfate (C6S), chondroitin (COS) and dermatan sulfate (DS) were identified by three monoclonal antibodies (2B6, 3B3 and 1B5) after chondroitinase ABC, B or AC II digestion. These antibodies recognized C4S, C6S, COS and DS stubs remaining on the core proteins of the PGs after enzyme digestion. The DS- and/or C6S-containing PGS were identified both in the E-and G2-extracts; the molecular masses of the core proteins of DS-PGs were approximately 45 kDa in both extracts, whereas two bands of the digested C6S-PGs (approximately 45 and 200 kDa) were present in the E-extract and only one C6S-PG band (approximately 200 kDa) was observed in the G2-extract. On the other hand, C4S- and COS-PGs were identified only in the G2-extract; the molecular mass of the digested C4S-PG was approximately 45 kDa, and that of the digested COS-PGs was approximately 200 kDa.
硬組織再生生物学会の論文