Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors
文献类型：期刊论文
| 作者 | Fan Cuiying2; Feng Lixing1; Zhang Dongmei1; Pan Suna2; Liu Xuan1; Guo Dean1 ; Fan Jinling2
|
| 刊名 | Chinese Journal of Biotechnology
 |
| 出版日期 | 2012 |
| 卷号 | 28期号:2页码:233-242 |
| 关键词 | PSMB1 BIAcore PSMB1 prokaryotic expression BIAcore screening in vitro |
| ISSN号 | 1000-3061 |
| 其他题名 | 人蛋白酶体亚基PSMB1的重组表达、纯化及在蛋白酶体抑制剂体外筛选中的应用 |
| 文献子类 | Article |
| 英文摘要 | Proteasome is a multi-subunit protease complex in eukaryocytes,and plays an important role in ubiquitin-proteosome pathway.Recombinant proteasome can be used to screen proteasome inhibitors.In this study,recombinant plasmid of pET28a-PSMB1 was constructed by inserting human proteasome catalytic subunit(PSMB1) cDNA(726 bp) into the prokaryotic expression vector pET28a(+),and transforming the plasmid into E.coli BL21(DE3) cells for expression.After overnight induction(1 mmol/L IPTG,20 °C),an expected protein band with molecular weight of 27 kDa was observed on SDS-PAGE gel.The recombinant protein was then purified through affinity chromatography,and the purity is more than 95%.The amino acid sequence of the recombinant protein was validated by NanoLC-MS/MS.The data from in vitro BIAcore analysis showed that the recombinant PSMB1 could bind to celastrol.The binding affinity between PSMB1 and 10 mumol/L celastrol was more than 27RU. |
| WOS研究方向 | Pharmacology & Pharmacy (provided by Clarivate Analytics) |
| 语种 | 中文 |
| CSCD记录号 | CSCD:4459689 |
| 源URL | [http://119.78.100.183/handle/2S10ELR8/267850]  |
| 专题 | 上海中药现代化研究中心 |
| 通讯作者 | Guo Dean |
| 作者单位 | 1.Shanghai Institute of Materia Medica,Chinese Academy of Sciences, Shanghai 201203, China. 2.College of Food & Engineering,Henan University of Science and Technology, Luoyang, Henan 471003, China.; |
| 推荐引用方式 GB/T 7714 | Fan Cuiying,Feng Lixing,Zhang Dongmei,et al. Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors[J]. Chinese Journal of Biotechnology,2012,28(2):233-242. |
| APA | Fan Cuiying.,Feng Lixing.,Zhang Dongmei.,Pan Suna.,Liu Xuan.,...&Fan Jinling.(2012).Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors.Chinese Journal of Biotechnology,28(2),233-242. |
| MLA | Fan Cuiying,et al."Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors".Chinese Journal of Biotechnology 28.2(2012):233-242. |
入库方式： OAI收割
来源：上海药物研究所
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