Evidence of block and randomly sequenced chondroitin polysaccharides: Sequential enzymatic digestion and quantification using ion trap tandem mass spectrometry
Evidence of block and randomly sequenced chondroitin polysaccharides: Sequential enzymatic digestion and quantification using ion trap tandem mass spectrometry
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DOI：
10.1021/ac010385j
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发表时间：
2001-08-01
影响因子：
7.4
通讯作者：
Leary, JA
中科院分区：
文献类型：
--
作者：
Desaire, H;Sirich, TL;Leary, JA
介绍了一种测定软骨样品中二糖重复序列类型的方法。将样品依次进行选择性和非选择性酶消化，并使用串联质谱法对每个步骤的异构体产物进行定量。两步消化/定量方案鉴定聚合物的总体组成相对于软骨的两种主要组分4-和6-硫酸化二糖是“交替的”、“随机的”还是“封闭的”。使用此程序，两个生物分离的软骨素多糖的序列类型进行了鉴定。从牛气管分离的硫酸软骨素A的结果与随机分布在多糖的整个重复区域的4-和6-硫酸化二糖一致。对于硫酸软骨素C，鲨鱼软骨，6-硫酸化二糖彼此相邻的程度比随机分布的聚合物预期的程度更大，表明存在具有相同硫酸化位点的重复二糖的“嵌段”。
A method for determining the sequence type of the disaccharide repeat region of cartilage samples is introduced. The samples are sequentially subjected to selective and nonselective enzymatic digestion, and the isomeric products from each step are quantified using tandem mass spectrometry. The two-step digestion/quantification protocol identifies whether the global makeup of the polymer is "alternating", "random", or "blocked" with respect to the two main components of the cartilage, 4-and 6-sulfated disaccharides. Using this procedure, the sequence type of two biologically isolated chondroitin polysaccharides was identified. The results for chondroitin sulfate A, isolated from bovine trachea, are consistent with the 4- and 6-sulfated disaccharides randomly distributed throughout the repeat region of the polysaccharide. For chondroitin sulfate C, shark cartilage, the 6-sulfated disaccharides are adjacent to each other to a larger extent than one would expect for a randomly distributed polymer, indicating that "blocks" of repeating disaccharides with the same sulfation site are present.