title: High-throughput system for analyzing ligand-induced cofactor recruitment by vitamin D receptor.
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.
authors.affiliation: Faculty of Pharmaceutical Sciences, Teikyo University, Sagamiko, Kanagawa 199-0195, Japan.
authors.forename: Midori A
authors.initials: MA
authors.lastname: Arai
authors.forename: Ken-ichi
authors.initials: K
authors.lastname: Takeyama
authors.forename: Saya
authors.initials: S
authors.lastname: Ito
authors.forename: Shigeaki
authors.initials: S
authors.lastname: Kato
authors.forename: Tai C
authors.initials: TC
authors.lastname: Chen
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kittaka
mesh_terms: D002232:Carbocyanines / Q000032:analysis; D045744:Cell Line, Tumor; D003065:Coenzyme A / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D005557:Formaldehyde / Q000737:chemistry; D051548:Histone Acetyltransferases / Q000032:analysis / Q000378:metabolism; D006801:Humans; D008024:Ligands; D056920:Nuclear Receptor Coactivator 1; D051958:Nuclear Receptor Coactivator 2 / Q000032:analysis / Q000378:metabolism; D018167:Receptors, Calcitriol / Q000819:agonists* / Q000378:metabolism; D014157:Transcription Factors / Q000032:analysis / Q000378:metabolism; D014807:Vitamin D / Q000031:analogs & derivatives* / Q000494:pharmacology*
chemical_list: D002232:Carbocyanines; D008024:Ligands; C498001:NCOA2 protein, human; D051958:Nuclear Receptor Coactivator 2; D018167:Receptors, Calcitriol; D014157:Transcription Factors; C089187:cyanine dye 3; C085321:cyanine dye 5; D014807:Vitamin D; D005557:Formaldehyde; D051548:Histone Acetyltransferases; C538859:NCOA1 protein, human; D056920:Nuclear Receptor Coactivator 1; D003065:Coenzyme A