Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors

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	Expression,purification of poteasome subunit PSMB1 and application in screening of possible proteasome inhibitors
	Fan Cuiying 2; Feng Lixing 1; Zhang Dongmei 1; Pan Suna 2; Liu Xuan 1; Guo Dean1 ; Fan Jinling 2
刊名	Chinese Journal of Biotechnology
	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
内容类型	期刊论文
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.