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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
1
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pubmed:dateCreated |
2005-12-13
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pubmed:abstractText |
The 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.
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Carbon Isotopes,
http://linkedlifedata.com/resource/pubmed/chemical/DNA-Binding Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Ligands,
http://linkedlifedata.com/resource/pubmed/chemical/NR5A2 protein, human,
http://linkedlifedata.com/resource/pubmed/chemical/Nitrogen Isotopes,
http://linkedlifedata.com/resource/pubmed/chemical/Receptors, Cytoplasmic and Nuclear,
http://linkedlifedata.com/resource/pubmed/chemical/Recombinant Proteins,
http://linkedlifedata.com/resource/pubmed/chemical/Trans-Activators,
http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors
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pubmed:status |
MEDLINE
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pubmed:month |
Jan
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pubmed:issn |
1046-5928
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
45
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
99-106
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pubmed:dateRevised |
2006-11-15
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pubmed:meshHeading |
pubmed-meshheading:16039139-Amino Acid Sequence,
pubmed-meshheading:16039139-Carbon Isotopes,
pubmed-meshheading:16039139-Circular Dichroism,
pubmed-meshheading:16039139-DNA-Binding Proteins,
pubmed-meshheading:16039139-Humans,
pubmed-meshheading:16039139-Isotope Labeling,
pubmed-meshheading:16039139-Ligands,
pubmed-meshheading:16039139-Light,
pubmed-meshheading:16039139-Magnetic Resonance Spectroscopy,
pubmed-meshheading:16039139-Molecular Sequence Data,
pubmed-meshheading:16039139-Nitrogen Isotopes,
pubmed-meshheading:16039139-Protein Structure, Secondary,
pubmed-meshheading:16039139-Protein Structure, Tertiary,
pubmed-meshheading:16039139-Receptors, Cytoplasmic and Nuclear,
pubmed-meshheading:16039139-Recombinant Proteins,
pubmed-meshheading:16039139-Scattering, Radiation,
pubmed-meshheading:16039139-Spectrometry, Mass, Electrospray Ionization,
pubmed-meshheading:16039139-Trans-Activators,
pubmed-meshheading:16039139-Transcription Factors
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pubmed:year |
2006
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pubmed:articleTitle |
Over-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.
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pubmed:affiliation |
State 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.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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