Efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith
文献类型：期刊论文
| 作者 | Ma, Junfeng1,2; Hou, Chunyan1,2; Liang, Yu1,2; Wang, Tingting1,2; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1 |
| 刊名 | proteomics
 |
| 出版日期 | 2011-03-01 |
| 卷号 | 11期号:5页码:991-995 |
| 关键词 | Immobilized enzyme reactor Metal-ion chelation Organic-inorganic hybrid silica monolith Protein digestion Technology Trypsin |
| ISSN号 | 待补充 |
| 产权排序 | 1,1 |
| 通讯作者 | 张丽华 |
| 中文摘要 | efficient proteolysis using a regenerable metal-ion chelate immobilized enzyme reactor supported on organic-inorganic hybrid silica monolith |
| 英文摘要 | 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. |
| WOS标题词 | science & technology ; life sciences & biomedicine |
| 学科主题 | 物理化学 |
| 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 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000288137300015 |
| 公开日期 | 2012-07-09 |
| 源URL | [http://159.226.238.44/handle/321008/115314]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 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 |
| 推荐引用方式 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.,...&Zhang, Yukui.(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. |
入库方式： OAI收割
来源：大连化学物理研究所
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