Lemperg R, Larsson S E, Jones I
Biochim Biophys Acta. 1977 Feb 28;496(2):420-30. doi: 10.1016/0304-4165(77)90324-5.
Articular cartilage from cow and calf femoral condyles was incubated in Tyrodes solution containing [35S]sulphate for different periods up to 80 min. Glycosaminoglycans from the cartilage tissue and incubation medium were fractionated on Cetylpyridinium chloride and ECTEOLA cellulose microcolumns. The incorporation of [35S]sulphate into all individual fractions of chondroitin sulphate and keratan sulphate was found to be linear from 20 to 80 min incubation time. As a rule the total specific activities of keratan sulphate and chondroitin sulphate were similar for both calves and cows. The proteoglycan material recovered from the medium amounted to about 1% of the tissue dry weight and was found to have a higher chondroitin sulphate: keratan sulphate ratio than the corresponding cartilage tissue for both calf and cow. The solubility profiles for the newly synthesised glycosaminoglycans, obtained from determination of the radioactivity in the individual fractions, were compared with those of glycosaminoglycans already present. These curves indicated that newly synthesised chondroitin sulphate had a higher average molecular size than that present in the tissue whereas the newly synthesised keratan sulphate had a smaller average molecular size. These newly synthesised components were also detected in the proteoglycans recovered from the incubation medium.
将来自牛和小牛股骨髁的关节软骨在含有[35S]硫酸盐的台氏液中孵育不同时间段，最长80分钟。软骨组织和孵育培养基中的糖胺聚糖在十六烷基氯化吡啶和ECTEOLA纤维素微柱上进行分级分离。发现在20至80分钟的孵育时间内，[35S]硫酸盐掺入硫酸软骨素和硫酸角质素的所有单个级分中的过程呈线性。通常，小牛和牛的硫酸角质素和硫酸软骨素的总比活性相似。从培养基中回收的蛋白聚糖物质约占组织干重的1%，并且发现其硫酸软骨素：硫酸角质素的比例高于小牛和牛相应的软骨组织。通过测定各个级分中的放射性获得的新合成糖胺聚糖的溶解度曲线与已存在的糖胺聚糖的溶解度曲线进行了比较。这些曲线表明，新合成的硫酸软骨素的平均分子大小比组织中存在的硫酸软骨素的平均分子大小更高，而新合成的硫酸角质素的平均分子大小更小。在从孵育培养基中回收的蛋白聚糖中也检测到了这些新合成的成分。
