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学科主题物理化学
Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith
Ma, Junfeng1,2; Hou, Chunyan1,2; Liang, Yu1,2; Wang, Tingting1,2; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1; Zhang LH(张丽华)
关键词Immobilized Enzyme Reactor Metal-ion Chelation Organic-inorganic Hybrid Silica Monolith Protein Digestion Technology Trypsin
刊名PROTEOMICS
2011-03-01
ISSN待补充
DOI10.1002/pmic.201000550
11期:5页:991-995
收录类别SCI
文章类型Article
部门归属18
项目归属1810
产权排名1,1
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods ; Biochemistry & Molecular Biology
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]MASS-SPECTROMETRY ; CAPILLARY MICROREACTOR ; PROTEIN IDENTIFICATION ; SOL-GEL ; TRYPSIN ; ONLINE ; ELECTROPHORESIS ; MICROSPHERES ; SEPARATION ; DIGESTION
英文摘要Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith; A metal-ion chelate immobilized enzyme reactor (IMER) supported on organic-inorganic hybrid silica monolith was developed for rapid digestion of proteins. The monolithic support was in situ prepared in a fused silica capillary via the polycondensation between tetraethoxysilane hydrolytic sol and iminodiacetic acid conjugated glycidoxypropyltrimethoxysilane. After activated by Cu(2+), trypsin was immobilized onto the monolithic support via metal chelation. Proteolytic capability of such an IMER was evaluated by the digestion of myoglobin and BSA, and the digests were further analyzed by microflow reversed-phase liquid chromatography with ESI-MS/MS. Similar sequence coverages of myoglobin and BSA were obtained by IMER, in comparison to those obtained by in-solution digestion (91 versus 92% for 200 ng myoglobin, and 26 versus 26% for 200 ng BSA). However, the digestion time was shortened from 12 h to 50 s. When the enzymatic activity was decreased after seven runs, the IMER could be easily regenerated by removing Cu(2+) via EDTA followed by trypsin immobilization with fresh Cu(2+) introduced, yielding the equal sequence coverage (26% for 200 ng BSA). For similar to 5 mu g rat liver extract, even more proteins were identified with the immobilized trypsin digestion within 150 s in comparison to the in-solution digestion for 24 h (541 versus 483), demonstrating that the IMER could be a promising tool for efficient and high-throughput proteome profiling.
语种英语
WOS记录号WOS:000288137300015
引用统计
被引频次:36[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/115314
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing, Peoples R China
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GB/T 7714
Ma, Junfeng,Hou, Chunyan,Liang, Yu,et al. Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith[J]. PROTEOMICS,2011,11(5):991-995.
APA Ma, Junfeng.,Hou, Chunyan.,Liang, Yu.,Wang, Tingting.,Liang, Zhen.,...&张丽华.(2011).Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith.PROTEOMICS,11(5),991-995.
MLA Ma, Junfeng,et al."Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith".PROTEOMICS 11.5(2011):991-995.
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