Studies on the Composition and Structure of Whale Cartilage Keratosulfate
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概要
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From whale nasal cartilage, keratosulfate barium salt was obtained in good yield by ethanol fractionation on cellulose column. The keratosulfate was further fractionated by anion exchange (Dowex 1) column chromatography. All the fractions obtained from the Dowex 1 column contained the various amounts of galactosamine, sialic acid, mannose, fucose and protein, in addition to the constituents ; glucosamine, galactose and sulfate. The ratios of sulfate to hexosamine in each fraction differed from 0.75 to 1.8. As sugar sulfates, galactose 6-sulfate other than N-acetylglucosamine 6-sulfate was detected in partial acid hydrolyzate by electrophoresis and by paper chromatography. The high content (11.7%) of sialic acid was remarkable in contrast with corneal keratosulfate in which the content of sialic acid was less than 1%. The sialic acid was isolated from whale cartilage keratosulfate and identified to be N-acetylneuraminic acid by paper chromatography, color reactions and infrared spectrum. The results of pronase and alkali treatments suggest that the sialic acid may be present in the carbohydrate moiety and not in the alkali labile peptide moiety of the keratosulfate. The sialic acid was easily released by neuraminidase digestion, which indicates that the sialic acid attaches to the non-reducing end of carbohydrate. In connection with sialic acid in cartilage, the sialic acid contents of cartilages from various sources were determined. The linkage between carbohydrate and peptide in the keratosulfate fraction was investigated by the action of alkali in the presence and absence of sodium borohydride. N-acetylhexosamine seemed to link to threonine and serine, and N-acetylhexosamine was assumed to be N-acetylgalactosamine.
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