Abstract
A high-throughput screening system for analyzing small molecule-induced coactivator (CoA) recruitment by the vitamin D receptor (VDR) has been developed. The vitamin D-induced protein-protein interactions between VDR and fluorophore (Cy3 or Cy5)-labeled TIF2 or SRC-1 were successfully detected by using a new HCHO fixing method of the protein complex on microplates. The results obtained from this screening of our synthetic vitamin D analogues suggest that the CoA-recruiting activities play an important role in determining the biological activity of various vitamin D analogues and explain the discrepancies between the VDR binding affinity and their biological activity.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
MeSH terms
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Carbocyanines / analysis
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Cell Line, Tumor
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Coenzyme A / metabolism*
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Drug Evaluation, Preclinical / methods*
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Formaldehyde / chemistry
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Histone Acetyltransferases / analysis
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Histone Acetyltransferases / metabolism
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Humans
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Ligands
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Nuclear Receptor Coactivator 1
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Nuclear Receptor Coactivator 2 / analysis
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Nuclear Receptor Coactivator 2 / metabolism
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Receptors, Calcitriol / agonists*
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Receptors, Calcitriol / metabolism
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Transcription Factors / analysis
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Transcription Factors / metabolism
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Vitamin D / analogs & derivatives*
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Vitamin D / pharmacology*
Substances
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Carbocyanines
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Ligands
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NCOA2 protein, human
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Nuclear Receptor Coactivator 2
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Receptors, Calcitriol
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Transcription Factors
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cyanine dye 3
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cyanine dye 5
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Vitamin D
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Formaldehyde
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Histone Acetyltransferases
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NCOA1 protein, human
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Nuclear Receptor Coactivator 1
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Coenzyme A