Novel screening system for high-affinity ligand of heredity vitamin D-resistant rickets-associated vitamin D receptor mutant R274L using bioluminescent sensor

Hiroki Mano, Miyu Nishikawa, Kaori Yasuda, Shinichi Ikushiro, Nozomi Saito, Daisuke Sawada, Shinobu Honzawa, Masashi Takano, Atsushi Kittaka, Toshiyuki Sakaki

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Hereditary vitamin D-resistant rickets (HVDRR) is caused by mutations in the vitamin D receptor (VDR) gene. Arg274 located in the ligand binding domain (LBD) of VDR is responsible for anchoring 1α,25-dihydroxyvitamin D3 (1α,25(OH)2D3) by forming a hydrogen bond with the 1α-hydroxyl group of 1α,25(OH)2D3. The Arg274Leu (R274L) mutation identified in patients with HVDRR causes a 1000-fold decrease in the affinity for 1α,25(OH)2D3, and dramatically reduces vitamin D- related gene expression. Recently, we successfully constructed fusion proteins consisting of split-luciferase and LBD of the VDR. The chimeric protein LucC-LBD-LucN, which displays the C-terminal domain of luciferase (LucC) at its N-terminus, can detect and discriminate between VDR agonists and antagonists. The LucC-LBD (R274L)-LucN was constructed to screen high-affinity ligands for the mutant VDR (R274L). Of the 33 vitamin D analogs, 5 showed much higher affinities for the mutant VDR (R274L) than 1α,25(OH)2D3, and 2α-[2-(tetrazol-2-yl)ethyl]-1α,25-(OH)2D3 showed the highest affinity. These compounds might be potential therapeutics for HVDRR caused by the mutant VDR (R274L).

Original languageEnglish
Pages (from-to)61-66
Number of pages6
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume167
DOIs
Publication statusPublished - Mar 1 2017

Fingerprint

Hypophosphatemic Rickets
Calcitriol Receptors
Heredity
Vitamin D
Screening
Ligands
Sensors
Luciferases
Mutation
Calcitriol
Gene expression
Hydroxyl Radical
Hydrogen
Hydrogen bonds
Proteins
Fusion reactions
Genes
Gene Expression

Keywords

  • Bioluminescent sensor
  • HVDRR
  • Rickets
  • Split luciferase
  • Vitamin D receptor

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Medicine(all)
  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Endocrinology
  • Clinical Biochemistry
  • Cell Biology

Cite this

Novel screening system for high-affinity ligand of heredity vitamin D-resistant rickets-associated vitamin D receptor mutant R274L using bioluminescent sensor. / Mano, Hiroki; Nishikawa, Miyu; Yasuda, Kaori; Ikushiro, Shinichi; Saito, Nozomi; Sawada, Daisuke; Honzawa, Shinobu; Takano, Masashi; Kittaka, Atsushi; Sakaki, Toshiyuki.

In: Journal of Steroid Biochemistry and Molecular Biology, Vol. 167, 01.03.2017, p. 61-66.

Research output: Contribution to journalArticle

Mano, Hiroki ; Nishikawa, Miyu ; Yasuda, Kaori ; Ikushiro, Shinichi ; Saito, Nozomi ; Sawada, Daisuke ; Honzawa, Shinobu ; Takano, Masashi ; Kittaka, Atsushi ; Sakaki, Toshiyuki. / Novel screening system for high-affinity ligand of heredity vitamin D-resistant rickets-associated vitamin D receptor mutant R274L using bioluminescent sensor. In: Journal of Steroid Biochemistry and Molecular Biology. 2017 ; Vol. 167. pp. 61-66.
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AU - Ikushiro, Shinichi

AU - Saito, Nozomi

AU - Sawada, Daisuke

AU - Honzawa, Shinobu

AU - Takano, Masashi

AU - Kittaka, Atsushi

AU - Sakaki, Toshiyuki

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