Glycosaminoglycans in Human and Bovine Serum: Detection of Twenty-Four Heparan Sulfate and Chondroitin Sulfate Motifs Including a Novel Sialic Acid-modified Chondroitin Sulfate Linkage Hexasaccharide.
Glycosaminoglycans in Human and Bovine Serum: Detection of Twenty-Four Heparan Sulfate and Chondroitin Sulfate Motifs Including a Novel Sialic Acid-modified Chondroitin Sulfate Linkage Hexasaccharide.
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DOI：
10.4137/gbi.s4273
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发表时间：
2010-02
期刊：
影响因子：
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通讯作者：
Hong Lu;L. McDowell;D. Studelska;Lijuan Zhang
中科院分区：
文献类型：
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作者：
Hong Lu;L. McDowell;D. Studelska;Lijuan Zhang
异质硫酸乙酰肝素和硫酸软骨素糖胺聚糖（GAG）多糖是血液循环的重要成分。癌症和其他人类疾病表明血液中 GAG 数量和结构的变化。然而，由于缺乏稳健和灵敏的分析工具，GAG 的数量和结构尚未得到充分表征。为了开发此类工具，我们从血清和血浆中分离了 GAG。我们采用液相色谱 (LC) 进行 GAG 定量，并采用 LC/质谱 (MS) 进行 GAG 结构分析。鉴定了二十四个乙酰肝素和硫酸软骨素基序，包括连接六糖、重复二糖组合物、还原端和非还原端单糖、二糖、三糖和四糖结构。无需放射性标记或衍生化即可在皮摩尔水平检测到二糖，因此本研究仅需要几毫升人和胎牛血清。与GAG连接六糖不同的不同还原末端结构的检测表明，GAG降解酶产生的游离GAG链与血清中的蛋白聚糖共存。此外，还发现一种新型唾液酸修饰的连接六糖与最丰富的血清蛋白聚糖 Bikunin 缀合。
Heterogeneous heparan sulfate and chondroitin sulfate glycosaminoglycan (GAG) polysaccharides are important components of blood circulation. Changes in GAG quantity and structure in blood have been indicated in cancers and other human diseases. However, GAG quantities and structures have not been fully characterized due to lack of robust and sensitive analytical tools. To develop such tools, we isolated GAGs from serum and plasma. We employed liquid chromatography (LC) for GAG quantification and LC/mass spectrometry (MS) for GAG structural analysis. Twenty-four heparan and chondroitin sulfate motifs were identified, including linkage hexasaccharides, repeating disaccharide compositions, reducing, and non-reducing end mono-, di-, tri-, and tetrasaccharide structures. Disaccharides were detectable at picomolar level without radiolabeling or derivitization, so only a few ml of human and fetal bovine serum was required for this study. The detection of different reducing end structures distinct from GAG linkage hexasaccharides revealed that free GAG chains generated by GAG degradation enzymes co-existed with proteoglycans in serum. In addition, a novel sialic acid-modified linkage hexasaccharide was found conjugated to bikunin, the most abundant serum proteoglycan.