高等学校化学学报

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小型微波谐振腔用于蛋白质微波辅助酶解

窦文超, 郇延富, 张志权, 张华容, 王超, 李明, 冯国栋, 金钦汉   

  1. 吉林省光谱分析仪器工程技术研究中心, 吉林大学化学学院, 长春 130023
  • 收稿日期:2006-06-21 修回日期:1900-01-01 出版日期:2007-02-10 发布日期:2007-02-10
  • 通讯作者: 冯国栋

Mini Microwave Resonance Cavity Used for Microwave-accelerated Protein-enzymolysis in Protein Mapping by Mass Spectrometry

DOU Wen-Chao, HUAN Yan-Fu, ZHANG Zhi-Quan, ZHANG Hua-Rong, WANG Chao, LI Ming, FENG Guo-Dong, JIN Qin-Han   

  1. Jilin Province Research Center for Engineering and Technology of Spectral Instruments, College of Chemistry, Jilin University, Changchun 130021, China
  • Received:2006-06-21 Revised:1900-01-01 Online:2007-02-10 Published:2007-02-10
  • Contact: FENG Guo-Dong

摘要: 采用微波谐振腔对细胞色素c以及牛血清白蛋白进行微波辅助酶解, 通过电喷雾三级四极杆质谱对得到的肽段进行分析, 证明该方法可用很低的微波功率将蛋白质彻底酶解为多肽. 通过调整微波条件可以使蛋白质的酶解效率基本达到100%, 细胞色素c和牛血清白蛋白的序列覆盖率分别为45%和26%. 该方法不但可将蛋白酶解时间由传统方法的16 h缩短为20 min, 还将功率由使用微波炉时的数百瓦降至20 W.

关键词: 微波谐振腔, 微波辅助酶解, 质谱, 选择性酶切

Abstract: A mini microwave resonance cavity was used to accelerate proteolytic cleavage of cytochrome c and bovine serum albumin. The resulting mixture were then analyzed by electrospray ionization mass spectrometer(ESI-MS).The digestion efficiencies of proteins was 100%,the sequence coverages of cytochrome c and bovine serum albumin were 45% and 26%, respectively. This approach not only speeds up the digestion process to 20 minutes versus 16 h by using traditional methods, but also reduced the power of microwave to 20 W versus hundreds Watt by using other microwave-assisted enzymatic digest methods.

Key words: Microwave resonance cavity, Microwave-accelerated protein digestion, Mass spectrometry, Selective endoprotease reactions

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