高等学校化学学报

高等学校化学学报 ›› 1999, Vol. 20 ›› Issue (6): 852.

• 论文 • 上一篇    下一篇

MALDI方法研究蛋白质的非共价复合物

佘益民1, 季怡萍2   

  1. 1. 湖南师范大学分析测试中心, 长沙 410006;
    2. 中国科学院长春应用化学研究所, 长春 130022
  • 收稿日期:1998-12-01 出版日期:1999-06-24 发布日期:1999-06-24
  • 通讯作者: 季怡萍
  • 作者简介:佘益民,男,33岁,博士,副教授,现在加拿大Manitoba大学.
  • 基金资助:

    湖南师范大学博士研究基金

Studies of Non-covalent Protein Complexes by MALDI Methods

SHE Yi-Min1, JI Yi-Ping2   

  1. 1. Analysis & Testing Center, Hunan Normal University, Changsha, 410006;
    2. Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022
  • Received:1998-12-01 Online:1999-06-24 Published:1999-06-24

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被引次数

6

摘要： 飞行时间质谱仪(TOFMS)在理论上无质量范围的限制,可实现大分子蛋白质与核酸的非共价复合物的直接检测.特别是在近中性溶液条件下通过对芥子酸和6-氮杂-2-硫代胸腺嘧啶基质的使用及双层样品制备方法的改善,获得了稳定复合物的高灵敏度质谱检测.肌红蛋白-血红素复合物能够在芥子酸基质的不同pH条件下(pH2.0或pH5.0)同时观察到.而运用双层样品制备方法,获得了核糖核酸酶复合物(RNaseS)在第一次激光照射下的突出质谱峰,但其丰度均随更多的激光打击而减弱.

关键词: 非共价复合物, 非共价蛋白质相互作用, 基质辅助解吸电离质谱

Abstract: Several specific non-covalent protein complexes were successfully observed by matrix assisted desorption ionization mass spectrometry(MALDI MS). The methods described in this paper include the matrixes use of sinapinic acid(SA) and 6-aza-2-thiothymine(ATT) in neutral pHsolution, as well as the improvement of two-layer sample preparation method to achieve a high sensitivity detection of stable non-covalent complexes. Myoglobin-heme complex was found simultaneously with the sinapinic acid matrix in the various pHsolution(pH=2 or pH=5). The RNase Scomplex showed a striking intensity at the first shot, which was decreased with more laser shots. Most importantly, the observation of specific noncovalent complex in the brome mosaic virus(BMV) coat proteins would open up a new possibility to investigate the assembly and disassembly of viral capsids.

Key words: Non-covalent complex, Non-covalent protein interaction, Matrix assisted desorption ionization mass spectrometry

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