高等学校化学学报

高等学校化学学报 ›› 2002, Vol. 23 ›› Issue (7): 1246.

• 研究论文 • 上一篇    下一篇

微柱高效液相色谱-质谱/质谱快速鉴定混合蛋白质新方法

张素艳1, 王静1, 卞利萍1, 许云敏2, 王洪海2,3, 杨芃原1,3   

  1. 1. 复旦大学化学系, 上海 200433;
    2. 复旦大学遗传研究所, 上海 200433;
    3. 蛋白质组学研究中心, 上海 200433
  • 收稿日期:2001-08-20 出版日期:2002-07-24 发布日期:2002-07-24
  • 通讯作者: 杨芃原(1949年出生),男,教授,博士生导师,从事生物质谱分析研究.E-mail:pyyang@fudan.edu.cn E-mail:pyyang@fudan.edu.cn
  • 基金资助:

    国家重点基础研究专项经费(批准号:2001CB5102);上海市科学技术委员会重点项目资助

A New Technique to Identify Individual Protein in Complex by Microcolumn-HPLC-ESIMS/MS

ZHANG Su-Yan1, WANG Jing1, BIAN Li-Ping1, XU Yun-Min2, WANG Hong-Hai2,3, YANG Peng-Yuan1,3   

  1. 1. Department of Chemistry, Shanghai 200433, China;
    2. Institute of Genetics, Shanghai 200433, China;
    3. Center of Proteomics, Fudan University, Shanghai 200433, China
  • Received:2001-08-20 Online:2002-07-24 Published:2002-07-24

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被引次数

37

摘要： 发展了一种混合蛋白质快速鉴定的新方法.将几种蛋白质的混合物于热变性后直接在溶液中酶解,利用微柱高效液相色谱-离子阱串级质谱进行肽谱/氨基酸序列分析,并结合Mascot数据库搜索处理功能,实现了混合蛋白质快速准确的鉴定.

关键词: 混合蛋白质鉴定, 蛋白质组学, 微柱高效液相色谱, 离子阱质谱/质谱, 序列分析

Abstract: In this work the identification method for protein mixture is reported, in which several proteins were varified by HPLC ESIMS/MSafter the whole complex proteins were directly digested in solution, instead of the routine 2-dimensional gel electrophoresis. The obtained data was then searched against NCBInr database by using MS/MS Ions Search(http://www.matrixscience.com), and a positive result was therefore obtained. With the simulated sequence spectra in database searching, a re identification method was developed for low-score matched protein by sequence-tag technique. As a typical example, Concanavalin A, a less-soluble protein with a low matching score, was also successfully identified, which indicated the great potential of complex digestion in proteomic study.

Key words: Identification of complex protein, Proteomics, Micro HPLC, Ion Trap tandem mass spectrometry, Sequence analysis

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