Carbohydrate-containing substances were extracted from non-calcified (NCC) and calcified (CC) portions of bovine costal cartilage with 0.5 M LaCl3 by the method of Mason and his co-workers, followed by dilution of the extract with 9 volumes of water. The precipitate formed on dilution yielded Fr. P, while Fr. S was obtained from the supernatant. Fr. P was separated into two subfractions by gel filtration on Sepharose 2B. The experimental results showed that Fr. P contained proteoglycans with different molecular sizes and compositions, while Fr. S contained proteoglycan, hyaluronic acid, glycoproteins, and glycogen. The present data suggest that in the proteoglycan of Fr. P, the relative content of chondroitin sulfate decreases with a concomitant increment in that of keratan sulfate on calcification. In addition, elevation of the ratio of chondroitin 4-sulfate to chondroitin 6-sulfate, together with a small increment of non-sulfated disaccharide units in the chondroitin sulfate chains appear to occur on calcification. The glycogen content in Fr. S diminished on calcification. The present observations suggest therefore that the remodeling of proteoglycan consumption of glycogen in bovine costal cartilage occur on calcification.
The supernatant fraction (Fr. S) obtained by ten-fold dilution of the 0.5 M LaCl3-extract of bovine costal cartilage was fractionated by DEAE-Sephadex A-25 column chromatography, followed by gel filtration on Sepharose 4B. The resulting five subfractions were characterized by electrophoresis on cellulose acetate membranes before and after digestion with pronase, x-amylase, and chondroitinase AC, together with analytical data. The results showed that Fr. S contained a large quantity of glycogen (Fr. 1), three sialoglycoproteins (Frs. 2, 3 and 4-1), hyaluronic acid (Fr. 4-1), a small molecular size proteoglycan consisting of chondroitin sulfates and keratan sulfate (Fr. 4-2), and small quantities of collagen-like protein.The components in the corresponding subfractions of Fr. S from non-calcified (NCC) and calcified (CC) portions of the costal cartilage were similar. It is indicated that consumption of glycogen is remarkable on calcification. glycan; glycogen; sialoglycoprotein; hyaluronic acid; proteoglycan; chondroitin sulfate; keratan sulfate; bovine costal cartilage In a previous paper (Futami et al. 1979), carbohydrate-containing substances were extracted with 0.5 M LaCl3 from noncalcified (NCC) and calcified (CC) portions of bovine costal cartilage, followed by dilution of the extract with 9 volumes of water. The precipitate (Fr. P) formed on dilution was separated into two subfractions by gel filtration on Sepharose 2B. On the other hand, Fr. S was obtained from the supernatant of the diluted LaCl3 solution after successive removal of the precipitates with 5 and 6% Na2SO4. Comparative studies on Fr. P, Fr. S and the subfractions of Fr. P from NCC and CC gave an important information on calcification of the cartilage.To obtain more precise information on the components in Fr. S, it was frac tionated by DEAE-Sephadex A-25 column chromatography, followed by gel filtra tion on Sepharose 4B. The resulting five subfractions from NCC and CC were characterized. This paper reports the results of the experiments.
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