| 猪肝组织中硫酸乙酰肝素的分离纯化及二糖组成分析 |
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| 引用本文： | 吕惠中,于广利,赵峡,张峥. 猪肝组织中硫酸乙酰肝素的分离纯化及二糖组成分析[J]. 中国生化药物杂志, 2007, 28(5): 297-300 |
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| 作者姓名： | 吕惠中 于广利 赵峡 张峥 |
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| 作者单位： | 1. 中国海洋大学,医药学院,糖科学与糖工程室,山东,青岛,266003
2. 青岛大学,医学院,生物化学与分子生物学教研室,山东,青岛,266021 |
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| 基金项目： | 国家重点基础研究发展计划(973计划) |
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| 摘 要： | 目的从少量猪肝组织中分离纯化硫酸乙酰肝素(HS)并进行二糖组成分析。方法分别采用胰蛋白酶及中性蛋白酶对猪肝组织进行酶解,再分别采用AG 50W-X2强阳离子交换色谱、Sephadex G-10凝胶过滤色谱、Q-Sepharose Fast Flow强阴离子交换色谱法纯化。将纯化的HS分别采用肝素酶Ⅰ和Ⅲ降解,所得到的二糖经2-氨基苯甲酰胺标记后,采用PA03色谱柱分析其二糖组成。结果经色谱和电泳分析,所得到的HS不含蛋白质及其它糖胺聚糖,其二糖组成以非硫酸化和低硫酸化二糖为主。结论该方法可以有效分离、纯化猪肝组织中的HS,并适用于少量组织中HS二糖组成分析。
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| 关 键 词： | 猪肝 硫酸乙酰肝素 纯化 二糖组成分析 |
| 文章编号： | 1005-1678（2007）05-0297-04 |
| 收稿时间： | 2007-02-05 |
| 修稿时间： | 2007-04-10 |
Isolation, purification and disaccharide composition analysis of heparan sulfate from porcine liver |
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| LU Hui-zhong,YU Guang-li,ZHAO Xia,ZHANG Zheng. Isolation, purification and disaccharide composition analysis of heparan sulfate from porcine liver[J]. Chinese Journal of Biochemical Pharmaceutics, 2007, 28(5): 297-300 |
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| Authors: | LU Hui-zhong YU Guang-li ZHAO Xia ZHANG Zheng |
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| Affiliation: | 1. Glycoscience and Glycoengineering Laboratory, Ocean University of China, Qingdao 266003, China ; 2. Department of Biochemistry and Molecular Biology, Medical School, Qingdao University, Qingdao 266021, China |
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| Abstract: | Purpose To isolate and to purify heparan sulfate(HS)from a small amount of porcine liver tissue and to analyze its disaccharide composition.Methods The HS was digested with trypsin and neutrase,and then purified by chromatography with AG 50W-X2,Sephadex G-10 and Q-Sepharose Fast Flow columns,respectively.The purified HS was treated with heparinase Ⅰand Ⅲ,the resulted disaccharides were labeled with 2-aminobenzamide(2-AB) and analyzed by HPLC on a PA03 column.Results The chromatography and electrophoresis data showed that the product was free of proteins and other glycosaminoglycans.Non-sulfated and low sulfated disaccharides were major components in porcine liver HS.Conclusion The method can be efficiently used to extract,purify and analyze disaccharide composition of HS from a small amount of liver tissue. |
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| Keywords: | porcine liver heparan sulfate purification disaccharide composition analysis |
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