Linked Life Data

16039139

Source:http://linkedlifedata.com/resource/pubmed/id/16039139

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pubmed-article:16039139pubmed:abstractTextThe human hepatitis B virus enhancer II B1 binding factor (hB1F), which regulates the expression of hepatitis B virus genes, is identified as a nuclear receptor. It regulates several liver-specific genes and plays an important role in the bile acid biosynthesis pathway. A significantly optimized protocol has been worked out to prepare 15N and/or 13C-labeled hB1F ligand-binding domain in minimal medium with high yields for NMR studies. Under the various conditions optimized for the purification of His6-hB1F ligand-binding domain, the yield of the purified protein is estimated to be 25-30 mg from 0.5 L of M9 minimal media. Electrospray ionization mass spectrometry data confirm the correctness of the primary sequence. Dynamic light scattering experiment proves that the protein exists as a monomeric form. In addition, the circular dichroism results show that the protein has a well-regulated secondary structure and a high alpha-helical content in ammonium bicarbonate buffer at 20 degrees C and pH 7.4. Finally, uniformly 15N-labeled protein is characterized by a TROSY-HSQC spectrum, and the dispersion of 15N-1H cross-peaks in the spectrum indicates the presence of well-ordered and properly folded protein as a monomer.lld:pubmed
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pubmed-article:16039139pubmed:authorpubmed-author:WangYuanYlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:ChenXiangXlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:WuHoumingHlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:MaJinbiaoJlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:ChenHaibaoHlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:XieYouhuaYlld:pubmed
pubmed-article:16039139pubmed:authorpubmed-author:TongXiaotianXlld:pubmed
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pubmed-article:16039139pubmed:dateRevised2006-11-15lld:pubmed
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pubmed-article:16039139pubmed:year2006lld:pubmed
pubmed-article:16039139pubmed:articleTitleOver-expression and purification of isotopically labeled recombinant ligand-binding domain of orphan nuclear receptor human B1-binding factor/human liver receptor homologue 1 for NMR studies.lld:pubmed
pubmed-article:16039139pubmed:affiliationState Key Laboratory of Bio-organic and Natural Products Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai 200032, People's Republic of China.lld:pubmed
pubmed-article:16039139pubmed:publicationTypeJournal Articlelld:pubmed
pubmed-article:16039139pubmed:publicationTypeResearch Support, Non-U.S. Gov'tlld:pubmed