Longas M O, Meyer K
Proc Natl Acad Sci U S A. 1982 Oct;79(20):6225-8. doi: 10.1073/pnas.79.20.6225.
Dermatan sulfate, purified by standard methods, displayed one spot at the position of the standard after two-dimensional cellulose acetate electrophoresis and was 99% in GalN, 21.5% in sulfate, and 0.6% in protein; Gal and Xyl (2:1) were the only neutral sugars detected. Its glucuronic acid/iduronic acid ratio was 0.15 and its Mr was approximately equal to 16,000. On reaction with 0.4 M NaOH, its reducing group(s) determined as Glc increased by 71% with concomitant separation of protein and polysaccharide and no alteration of the amino acids; when this reaction was repeated in the presence of 0.3 M NaBH4, only 31% of Lys was detected by standard amino acid analysis and none by TLC of the dansylated amino acids; alkaline cleavage in these conditions yielded only 30% of the original Xyl, xylitol, and a ninhydrin-positive substance different from GalN, which had the retention time of xylitol on Affi-Gel 601 and was also obtained from reduction of dermatan sulfate with a 400-eq excess of Na3BH4 in water under conditions that did not cleave the dermatan sulfate-protein bond. The data indicate that a reducible xylosyl-lysine is the protein linkage of dermatan sulfate from calf ligamentum nuchae.
经标准方法纯化的硫酸皮肤素，在二维醋酸纤维素电泳后在标准位置显示一个斑点，其GalN含量为99%，硫酸盐含量为21.5%，蛋白质含量为0.6%；检测到的唯一中性糖是Gal和Xyl（2:1）。其葡萄糖醛酸/艾杜糖醛酸比率为0.15，Mr约等于16,000。与0.4 M NaOH反应时，其确定为Glc的还原基团增加了71%，同时蛋白质和多糖分离，氨基酸无变化；当在0.3 M NaBH4存在下重复此反应时，通过标准氨基酸分析仅检测到31%的Lys，丹磺酰化氨基酸的TLC未检测到Lys；在这些条件下进行碱性裂解仅产生30%的原始Xyl、木糖醇和一种茚三酮阳性物质，该物质不同于GalN，在Affi-Gel 601上具有木糖醇的保留时间，并且也可通过在不裂解硫酸皮肤素-蛋白质键的条件下用400当量过量的Na3BH4在水中还原硫酸皮肤素来获得。数据表明可还原的木糖基赖氨酸是来自小牛项韧带的硫酸皮肤素的蛋白质连接键。
