IR@PKUHSC  > 北京大学第三临床医学院  > 心血管内科
学科主题临床医学
Reversible and oriented immobilization of histidine-tagged protein on silica gel characterized by frontal analysis
Gao, Xiaokang1; Li, Yunzhe1; Qin, Yu1; Chen, Enhui1; Li, Qian1; Zhao, Xinfeng1,2; Bian, Liujiao1; Zheng, Jianbin2; Li, Zijian3; Zhang, Youyi3; Zheng, Xiaohui1,2
刊名RSC ADVANCES
2015
DOI10.1039/c5ra01012h
5期:31页:24449-24454
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Chemistry, Multidisciplinary
研究领域[WOS]Chemistry
关键词[WOS]BETA(2)-ADRENOCEPTOR ; NANOPARTICLES ; CHROMATOGRAPHY ; ADRENOCEPTOR ; ORIENTATION ; INTERFACES ; DISCOVERY ; BETA(2) ; BINDING ; SCREEN
英文摘要

This approach utilized N,N′-bis(carboxymethyl)-L-lysine (ANTA) coordinated to bivalent metal cation Ni2+, leaving free coordination sites for the reversible binding of gene recombinant histidine-tagged beta(2)-adrenoceptor onto macropore silica. The amount of transient metal nickel ion on the support was determined by atomic absorption spectrophotometry. The novel protein oriented immobilization beta(2)-AR column was evaluated by five beta(2)-adrenoceptor agonists, applying frontal analysis. The association equilibrium constant for ligands on the column was 1.98 x 10(4) M-1 for salbutamol, 3.43 x 10(4) M-1 for clenbuterol, 2.09 x 10(4) M-1 for tulobuterol, 1.84 x 10(4) M-1 for terbutaline, 1.71 x 10(4) M-1 for methoxyphenamine and corresponding concentrations at binding sites were 7.46 x 10(-6) M, 1.82 x 10(-5) M, 2.16 x 10(-5) M, 8.29 x 10(-6) M and 3.88 x 10(-5) M, respectively. The results obtained from breakthrough and nonlinear fitting indicated that all the drugs have a single binding site on the beta(2)-adrenoceptor column. The present combined histidine-tagged protein method was reliable and exact in revealing interactions between receptor and drugs.

语种英语
WOS记录号WOS:000351236300050
项目编号21475103 ; 2013KCT-24 ; 2013YQ170525 ; 2013YQ17052509
资助机构National Nature Science Foundation of China ; program for Innovative Research Team of Shaanxi Province ; Ministry of Science and Technology of the People&prime ; s Republic of China
引用统计
被引频次:3[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/53661
专题北京大学第三临床医学院_心血管内科
作者单位1.NW Univ Xian, Minist Educ, Key Lab Resource Biol & Biotechnol Western China, Xian 710069, Peoples R China
2.NW Univ Xian, Inst Analyt Sci, Xian 710069, Peoples R China
3.Peking Univ, Hosp 3, Inst Vasc Med, Key Lab Mol Cardiovasc Sci,Minist Educ, Beijing 100083, Peoples R China
推荐引用方式
GB/T 7714
Gao, Xiaokang,Li, Yunzhe,Qin, Yu,et al. Reversible and oriented immobilization of histidine-tagged protein on silica gel characterized by frontal analysis[J]. RSC ADVANCES,2015,5(31):24449-24454.
APA Gao, Xiaokang.,Li, Yunzhe.,Qin, Yu.,Chen, Enhui.,Li, Qian.,...&Zheng, Xiaohui.(2015).Reversible and oriented immobilization of histidine-tagged protein on silica gel characterized by frontal analysis.RSC ADVANCES,5(31),24449-24454.
MLA Gao, Xiaokang,et al."Reversible and oriented immobilization of histidine-tagged protein on silica gel characterized by frontal analysis".RSC ADVANCES 5.31(2015):24449-24454.
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