PMID- 21397016
OWN - NLM
STAT- MEDLINE
DCOM- 20110801
LR  - 20240506
IS  - 1879-1220 (Electronic)
IS  - 0960-0760 (Print)
IS  - 0960-0760 (Linking)
VI  - 125
IP  - 3-5
DP  - 2011 Jul
TI  - A human vitamin D receptor mutant activated by cholecalciferol.
PG  - 202-10
LID - 10.1016/j.jsbmb.2011.03.001 [doi]
AB  - The human vitamin D receptor (hVDR) is a member of the nuclear receptor 
      superfamily, involved in calcium and phosphate homeostasis; hence implicated in a 
      number of diseases, such as Rickets and Osteoporosis. This receptor binds 
      1alpha,25-dihydroxyvitamin D(3) (also referred to as 1,25(OH)(2)D(3)) and other known 
      ligands, such as lithocholic acid. Specific interactions between the receptor and 
      ligand are crucial for the function and activation of this receptor, as implied 
      by the single point mutation, H305Q, causing symptoms of Type II Rickets. In this 
      work, further understanding of the significant and essential interactions between 
      the ligand and the receptor was deciphered, through a combination of rational and 
      random mutagenesis. A hVDR mutant, H305F, was engineered with increased 
      sensitivity towards lithocholic acid, with an EC(50) value of 10 muM and 40+/-14 
      fold activation in mammalian cell assays, while maintaining wild-type activity 
      with 1,25(OH)(2)D(3). Furthermore, via random mutagenesis, a hVDR mutant, 
      H305F/H397Y, was discovered to bind a novel small molecule, cholecalciferol, a 
      precursor in the 1alpha,25-dihydroxyvitamin D(3) biosynthetic pathway, which does not 
      activate wild-type hVDR. This variant, H305F/H397Y, binds and activates in 
      response to cholecalciferol concentrations as low as 100 nM, with an EC(50) value 
      of 300 nM and 70+/-11 fold activation in mammalian cell assays. In silico docking 
      analysis of the variant displays a dramatic conformational shift of 
      cholecalciferol in the ligand binding pocket in comparison to the docked analysis 
      of cholecalciferol with wild-type hVDR. This shift is hypothesized to be due to 
      the introduction of two bulkier residues, suggesting that the addition of these 
      bulkier residues introduces molecular interactions between the ligand and 
      receptor, leading to activation with cholecalciferol.
CI  - Copyright (c) 2011 Elsevier Ltd. All rights reserved.
FAU - Ousley, Amanda M
AU  - Ousley AM
AD  - School of Chemistry and Biochemistry, Parker H. Petit Institute for 
      Bioengineering and Biosciences, Georgia Institute of Technology, 901 Atlantic 
      Drive, Atlanta, GA 30332, United States. gth783d@mail.gatech.edu
FAU - Castillo, Hilda S
AU  - Castillo HS
FAU - Duraj-Thatte, Anna
AU  - Duraj-Thatte A
FAU - Doyle, Donald F
AU  - Doyle DF
FAU - Azizi, Bahareh
AU  - Azizi B
LA  - eng
GR  - R01 AI064817/AI/NIAID NIH HHS/United States
GR  - R01 AI064817-02/AI/NIAID NIH HHS/United States
GR  - 5R01AI064817-02/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20110310
PL  - England
TA  - J Steroid Biochem Mol Biol
JT  - The Journal of steroid biochemistry and molecular biology
JID - 9015483
RN  - 0 (Bone Density Conservation Agents)
RN  - 0 (Receptors, Calcitriol)
RN  - 1C6V77QF41 (Cholecalciferol)
SB  - IM
MH  - Bone Density Conservation Agents/*pharmacology
MH  - Cell Line
MH  - Cholecalciferol/*pharmacology
MH  - Gene Expression Regulation/*drug effects
MH  - Humans
MH  - Mutagenesis
MH  - Polymerase Chain Reaction
MH  - Protein Engineering/methods
MH  - Receptors, Calcitriol/chemistry/genetics/*metabolism
PMC - PMC3105188
MID - NIHMS285733
EDAT- 2011/03/15 06:00
MHDA- 2011/08/02 06:00
PMCR- 2012/07/01
CRDT- 2011/03/15 06:00
PHST- 2010/06/23 00:00 [received]
PHST- 2011/02/28 00:00 [revised]
PHST- 2011/03/04 00:00 [accepted]
PHST- 2011/03/15 06:00 [entrez]
PHST- 2011/03/15 06:00 [pubmed]
PHST- 2011/08/02 06:00 [medline]
PHST- 2012/07/01 00:00 [pmc-release]
AID - S0960-0760(11)00044-6 [pii]
AID - 10.1016/j.jsbmb.2011.03.001 [doi]
PST - ppublish
SO  - J Steroid Biochem Mol Biol. 2011 Jul;125(3-5):202-10. doi: 
      10.1016/j.jsbmb.2011.03.001. Epub 2011 Mar 10.