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化学学报
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化学学报 ›› 2011, Vol. 69 ›› Issue (18): 2123-2129. 上一篇    下一篇

研究论文

小尺度4-巯基苯硼酸修饰纳米金粒子用于大鼠肝脏糖蛋白选择性富集和质谱鉴定研究

仇娟，张莹，陆豪杰*，杨芃原*   

  1. (复旦大学化学系和生物医学研究院 上海 200032)
  • 投稿日期:2011-04-19 修回日期:2011-05-20 发布日期:2011-05-23
  • 通讯作者: 陆豪杰 E-mail:luhaojie@fudan.edu.cn
  • 基金资助:

    上海高校特聘教授(东方学者)岗位计划;上海市教育委员会和上海市教育发展基金会“曙光计划”;上海市重点学科;国家自然科学基金;国家自然科学基金;国家自然科学基金

Small Size 4-Mercaptophenylboronic Acid Functionalized Gold Nanoparticles for Selective Enrichment and Mass Spectrometric Identification of Glycoproteins in Rat Liver

CHOU Juan, ZHANG Ying, LU Hao-Jie, YANG Peng-Yuan   

  1. (Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200032)
  • Received:2011-04-19 Revised:2011-05-20 Published:2011-05-23

PDF

2056

被引次数

5 | 3

报道了利用硼氢化钠还原一步法合成小尺度巯基苯硼酸修饰的纳米金材料(尺寸约为3 nm), 合成方法简便快捷. 利用纳米金表面的硼酸基团与糖蛋白/糖肽中顺式邻二羟基的酯化反应, 成功实现了糖肽和糖蛋白的选择性富集. 利用该材料对糖蛋白HRP酶解产物进行富集, 经富集后糖肽质谱峰的信号强度提高近百倍|将该材料用于大鼠肝脏中糖蛋白的富集, 鉴定到了159158个N-糖基化位点对应于156个糖蛋白. 并对鉴定到的糖蛋白的分子功能进行了分析, 结果表明部分糖蛋白参与肝病相关的信号通路, 有着重要的生物学意义.

关键词: 巯基苯硼酸, 纳米金, 小尺度, 富集, 糖蛋白质, 质谱, 大鼠肝脏

In this work, small size 4-mercaptophenylboronic acid functionalized gold nanoparticles (the particle size was about 3 nm) were synthesized by a simple, one-step sodium borohydride reduction method. Based on the formation of diesters between boric acid group and cis-diol of glycoprotein/glycopeptieds, it was successfully applied to selectively enrich glycoprotein and glycopeptide. Using this material to enrich glycopeptides in the tryptic digests of HRP, the MS signals of glycopeptides were nearly hundredfold enhanced. The boronic acid functionalized nanoparticles were used to selectively capture of glycoprotein from rat liver. 1589 unique glycosylation sites mapped to 156 different glycoproteins have been identified and the molecular functions of these identified glycoproteins were analyzed. Results showed that some glycoproteins, for example Hspa8 and Shc1, played important biological roles related to liver disease.

Key words: 4-mercaptophenylboronic acid, gold nanoparticle, small size, enrichment, glycoprotein, mass spectrometry, rat liver

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