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共表达PXRLBD和SRC88以及PXR配体筛选的平衡透析模型的建立 被引量:1

Coexpression of PXRLBD with SRC88 and construction of equilibrium dialysis model of screening PXR ligands
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摘要 The aim of this study was to obtain the soluble protein of human pregnane X receptor ligand binding domain(PXRLBD) through the coexpression of PXRLBD and 88 amino acids of steroid receptor coactivator-1(SRC88) and apply the protein to constructing a new model of screening PXR ligands.Expression plasmid of pETDuet-1-SRC88-PXRLBD was constructed and transformed into Escherichia coli Rosetta(DE3) to coexpress PXRLBD and SRC88 via induction by IPTG at low temperature.Then an equilibrium dialysis model was constructed to study the interaction between PXRLBD and drugs including clotrimazole and dexamethasone,using HPLC as the analysis method.The results showed that the soluble protein of PXRLBD was obtained and the HPLC data indicated that clotrimazole bound to PXRLBD,while dexamethasone did not bind to PXRLBD,which indicated the successful establishment of a new method for studying the interaction between PXR and drugs.The new method may be useful in the screening of PXR ligands in vitro. The aim of this study was to obtain the soluble protein of human pregnane X receptor ligand binding domain (PXRLBD) through the coexpression of PXRLBD and 88 amino acids of steroid receptor coactivator-1 (SRC88) and apply the protein to constructing a new model of screening PXR ligands. Expression plasmid of pETDuet-l-SRC88-PXRLBD was constructed and transformed into Escherichia coli Rosetta (DE3) to coexpress PXRLBD and SRC88 via induction by IPTG at low temperature. Then an equilibrium dialysis model was constructed to study the interaction between PXRLBD and drugs including clotrimazole and dexamethasone, using HPLC as the analysis method. The results showed that the soluble protein of PXRLBD was obtained and the HPLC data indicated that clotrimazole bound to PXRLBD, while dexamethasone did not bind to PXRLBD, which indicated the successful establishment of a new method for studying the interaction between PXR and drugs. The new method may be useful in the screening of PXR ligands in vitro.
出处 《药学学报》 CAS CSCD 北大核心 2008年第4期427-430,共4页 Acta Pharmaceutica Sinica
基金 国家自然科学基金资助项目(30472171).
关键词 孕烷X受体 甾体受体辅活化因子 共表达 配体 透析 高效液相色谱法 PXR SRC coexpression ligand dialysis HPLC
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  • 1熊非,朱家壁,王维,华小兵.灯盏花素纳米脂质体包封率测定方法研究[J].药学学报,2004,39(9):755-757. 被引量:33
  • 2盛亮洪,李睿岩,李萍,邹汉法,孔亮.固定化脂质体色谱研究中药复方的细胞膜通透性成分及其质量控制[J].分析化学,2004,32(12):1595-1598. 被引量:20
  • 3盛亮洪,李萍,邹汉法.固定化脂质体色谱与动物小肠吸收模型的相关性研究方法及其应用[J].分析化学,2005,33(1):13-16. 被引量:16
  • 4David R. Cytochrome P450 and the individuality [J].Arch Biochem Biophy, 1999,369 : 1 - 10.
  • 5Kliewer SA, Moore JT, Wade L, et al. An orphan nuclear receptor activated by pregnanes defines a noval steroid signaling pathway [J]. Cell, 1998,92:73 - 82.
  • 6Watkins RE, Wisely GB, Moore LB, et al. The human nuclear xenobiotic receptor PXR: structural determinants of directed promiscuity [J]. Science, 2001,292:2329 -2333.
  • 7Xie W, Barwick JL, Downes M, et al. Humanized xenobiotic response in mice expressing nuclear receptor SXR [J]. Nature, 2000,406:435 - 439.
  • 8Zhang J, Kuehl P, Green ED, et al. The human pregnane X receptor:genomic structure and identification and functional characterization of natural allelic variants[J].Pharmacogenetics, 2001,11:555 - 572.
  • 9Phillips A, Hood SR, Gibson GG, et al. Impact of transcription factor profile and chromatin conformation on human hepatocyte CYP3A gene expression [J]. Drug Metab Dispos, 2005,33:233 - 242.
  • 10Song X, Xie M, Zhang H, et al. The pregnane X receptor binds to response elements in a genomic context-dependent manner and PXR activators rifampicin selectively alters the binding among target genes[J].Drug Metab Dispos, 2004,32:35 - 42.

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  • 1马俊锋,段继诚,梁振,张丽华,张维冰,张玉奎.固定化酶反应器及其在蛋白质组研究中的应用[J].分析化学,2006,34(11):1649-1655. 被引量:7
  • 2Lenz GR, Nash HM, Jindal S, et al. Chemical ligands, genomics and drug discovery [ J ]. Drug Discov Today, 2000,5 ( 4 ) : 145 - 156.
  • 3Stevenson DE, Feng R, Storer AC, et al. Detection of covalent enzyme-substrate complexes of nitrilase by ion-spray mass spectroscopy [ J ]. FEBS Lett, 1990,277 ( 1-2 ) : 112 - 114.
  • 4Baca M, Kent SBH. Direct observation of a ternary complex between the dimeric enzyme HIV-1 protease and a substratebased inhibitor[ J]. J Am Chem Soc, 1992,114 (10) :3 992- 3 993.
  • 5Stokvis E, Clugston SL, Honek JF, et al. Characterization of glyoxalase I (E. coli) -inhibitor interactions by electrospray time- of-flight mass spectrometry and enzyme kinetic analysis [ J ]. J Protein Chem,2000,19 ( 5 ) : 389 - 397.
  • 6Cheng XH, Chen R, Bruce JE ,et al. Using electrospray ionization FTICR mass spectrometry to study competitive binding of inhibitors to carbonic anhydrase [ J ] . J Am Chem Soc, 1995,117 (34) :8 859 -8 860.
  • 7Cummins LL, Chen S, Blyn LB, et al. Muhitarget affinity/specificity screening of natural products:finding and characterizing high-affinity ligands from complex mixtures by using high-performance mass spectrometry [ J ] . J Nat Prod, 2003,66 ( 9 ) : 1 186-1 190.
  • 8Griffey RH, Sannes-Lowery KA, Drader JJ, et al. Characterization of low-affinity complexes between RNA and small molecules using electrospray ionization mass spectrometry [ J ]. J Am Chem Soc,2000,122(41 ) :9 933-9 938.
  • 9Nikolic D, van Breemen RB. Screening for inhibitors of dihydrofolate reductase using pulsed ultrafihration mass spectrometry [ J ]. Comb Chem High Throughput Screen, 1998,1 ( 1 ) : 47 -55.
  • 10Nikolic D, Habibi-Goudarzi S, Corley DG, et al. Evaluation of cyclooxygenase-2 inhibitors using pulsed ultrafiltration mass spectrometry[J]. Anal Chem,2000,72(16) :3 853 -3 859.

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