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学科主题: 物理化学
题名: Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins
作者: Liu, Fangjie1, 2;  Ye, Mingliang1;  Wang, Chunli1, 2;  Hu, Zhengyan1, 2;  Zhang, Yi1, 2;  Qin, Hongqiang1, 2;  Cheng, Kai1, 2;  Zou, Hanfa1
通讯作者: 邹汉法 ;  叶明亮
刊名: ANALYTICAL CHEMISTRY
发表日期: 2013-08-06
DOI: 10.1021/ac4017693
卷: 85, 期:15, 页:7024-7028
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1809
产权排名: 1,1
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Trypsin was immobilized on a variety of materials to improve digestion efficiency. However, because the immobilized trypsin will digest proteins during electrophoresis, direct immobilization of active trypsin in polyacrylamide gel will compromise the protein separation. To overcome this problem, here we report a novel polyacrylamide gel with switchable trypsin activity. It was prepared by copolymerization of the PEG-trypsin-aprotinin complex during the gel-casting step. Because the inhibitor aprotinin binds strongly with trypsin at alkaline pH, this novel gel does not display hydrolytic activity during electrophoresis. After electrophoresis, the activity of trypsin embedded in gel could be recovered by simply washing away the bound inhibitor at a low pH. It was demonstrated that this unique switchable activity design allowed high resolution of the complex protein mixture during electrophoresis and highly efficient digestion of the separated proteins in situ in the gel after electrophoresis.
关键词[WOS]: MASS-SPECTROMETRY ;  IDENTIFICATION ;  PROTEOMICS ;  DIGESTION ;  MEMBRANES
语种: 英语
WOS记录号: WOS:000323014000009
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/117567
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China

Recommended Citation:
Liu, Fangjie,Ye, Mingliang,Wang, Chunli,et al. Polyacrylamide Gel with Switchable Trypsin Activity for Analysis of Proteins[J]. ANALYTICAL CHEMISTRY,2013,85(15):7024-7028.
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