HETEROGENEITY OF REDUCING TERMINALS OF URINARY CHONDROITIN SULFATES
HETEROGENEITY OF REDUCING TERMINALS OF URINARY CHONDROITIN SULFATES
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DOI：
10.1016/0304-4165(87)90056-0
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发表时间：
1987-03-19
期刊：
影响因子：
--
通讯作者：
ENDO, M
中科院分区：
文献类型：
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作者：
MATSUE, H;ENDO, M
用氯化十六烷基吡啶沉淀混合尿液中的不可透析部分，然后乙醇分级，然后用神经氨酸酶和粘多糖进行酶消化，分离出人尿软骨素硫酸盐。进一步用Dowex-1层析逐步洗脱，每隔0.25M将氯化钠浓度从0.75M增加到1.5M，得到的硫酸软骨素用糖、氨基酸和硫酸盐的分析和定量以及醋酸纤维素膜的电泳法进行了表征。用硼氢化钠还原还原末端后，用气-液层析法对水解物中的乙酰和丁基硼酸衍生物进行气液色谱分析，确定还原末端。在1.5M组分中，22.8%的尿硫酸软骨素是多肽结合的，其余的是无肽的，还原末端是木糖(29.8%)、半乳糖(23.6%)和葡萄糖醛酸(18.7%)。在0.75M-、1.0M-、1.25M-和1.5M-组分的还原末端含有木糖和半乳糖的无肽硫酸软骨素的量随硫酸盐化程度的增加和多肽含量的减少而增加。提示β-木糖苷酶、β-半乳糖苷酶和β-葡萄糖醛酸苷酶作用于该组织中蛋白-硫酸软骨素的糖-肽连接区，并在还原末端产生不同类型的尿硫酸软骨素。
Human urinary chondroitin sulfates were isolated by precipitation with cetylpyridinium chloride of the non-dialyzable fraction of pooled urine, followed by ethanol fractionation and successive enzymic digestions with neuraminidase and mucopolysaccharides. Further purification was achieved by Dowex-1 chromatography with stepwise elution by increasing the concentration of NaCl at intervals of 0.25 M from 0.75 M to 1.5 M. The chondroitin sulfates thus obtained were characterized by the analysis and quantification of carbohydrate, amino acid and sulfate, and by electrophoresis on cellulose acetate membrane. Then reducing terminals were identified by gas liquid chormatographic analyses of the acetyl and butaneboronate derivatives of hydrolysates, after reduction of the reducing terminals with sodium borohydride. About 22.8% of the urinary chondroitin sulfate in the 1.5 M fraction was peptide-bound, and the remainder was peptide-free, with xylose (29.8%), galactose (23.6%) and glucuronic acid (18.7%) at the reducing terminal. The amount of peptide-free chondrotinin sulfate with xylose and galactose at its reducing terminals in the 0.75 M-, 1.0 M-, 1.25 M- and 1.5 M-fractions increased in the order described in parallel with the increase of sulfation and the decrease of peptide content. It was thus suggested that the endo-types of .beta.-xylosidase, .beta.-galactosidase and .beta.-glucuronidase acted on the carbohydrate-peptide linkage region of proteo-chondroitin sulfate in this tissues and produced various types of urinary chondroitin sulfate with heterogeneity at reducing terminals.