TY - JOUR
T1 - Novel screening system for high-affinity ligand of heredity vitamin D-resistant rickets-associated vitamin D receptor mutant R274L using bioluminescent sensor
AU - Mano, Hiroki
AU - Nishikawa, Miyu
AU - Yasuda, Kaori
AU - Ikushiro, Shinichi
AU - Saito, Nozomi
AU - Sawada, Daisuke
AU - Honzawa, Shinobu
AU - Takano, Masashi
AU - Kittaka, Atsushi
AU - Sakaki, Toshiyuki
N1 - Funding Information:
This work was supported by Grants-in-Aid from the Japan Society for the Promotion of Science (No. 16H04912 and 16K14904 to T.S.)
Publisher Copyright:
© 2016 Elsevier Ltd
PY - 2017/3/1
Y1 - 2017/3/1
N2 - 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).
AB - 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).
KW - Bioluminescent sensor
KW - HVDRR
KW - Rickets
KW - Split luciferase
KW - Vitamin D receptor
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U2 - 10.1016/j.jsbmb.2016.11.008
DO - 10.1016/j.jsbmb.2016.11.008
M3 - Article
C2 - 27864003
AN - SCOPUS:85009267003
SN - 0960-0760
VL - 167
SP - 61
EP - 66
JO - Journal of Steroid Biochemistry
JF - Journal of Steroid Biochemistry
ER -
