PMID- 21884792
OWN - NLM
STAT- MEDLINE
DCOM- 20120210
LR  - 20211020
IS  - 1879-1220 (Electronic)
IS  - 0960-0760 (Linking)
VI  - 128
IP  - 1-2
DP  - 2012 Jan
TI  - The role of residue C410 on activation of the human vitamin D receptor by various 
      ligands.
PG  - 76-86
LID - 10.1016/j.jsbmb.2011.08.003 [doi]
AB  - Nuclear receptors (NRs) are ligand-activated transcription factors that regulate 
      the expression of genes involved in biologically important processes. The human 
      vitamin D receptor (hVDR) is a member of the NR superfamily and is responsible 
      for maintaining calcium and phosphate homeostasis. This receptor is activated by 
      its natural ligand, 1alpha, 25-dihydroxyvitamin D(3) (1alpha, 25(OH)(2)D(3)), as well as 
      bile acids such as lithocholic acid (LCA). Disruption of molecular interactions 
      between the hVDR and its natural ligand result in adverse diseases, such as 
      rickets, making this receptor a good target for drug discovery. Previous 
      mutational analyses of the hVDR have mainly focused on residues lining the 
      receptor's ligand binding pocket (LBP) and techniques such as alanine scanning 
      mutagenesis and site-directed mutagenesis. In this work, a rationally designed 
      hVDR library using randomized codons at selected positions provides insight into 
      the role of residue C410, particularly on activation of the receptor by various 
      ligands. A variant, C410Y, was engineered to bind LCA with increased sensitivity 
      (EC(50) value of 3 muM and a 34-fold activation) in mammalian cell culture assays. 
      Furthermore, this variant displayed activation with a novel small molecule, 
      cholecalciferol (chole) which does not activate the wild-type receptor, with an 
      EC(50) value of 4 muM and a 25-fold activation. The presence of a bulky residue at 
      this position, such as a tyrosine or phenylalanine, may contribute towards 
      molecular interactions that allow for the enhanced activation with LCA and novel 
      activation with chole. Additional bulk at the same end of the pocket, such as in 
      the case of the variant H305F; C410Y enhances the receptor's sensitivity for 
      these ligands further, perhaps due to the filling of a cavity. The effects of 
      residue C410 on specificity and activation with the different ligands studied 
      were unforeseen, as this residue does not line the hVDR's LBP. Further 
      investigating of the structure-function relationships between the hVDR and its 
      ligands, including the mutational tolerance of residues within as well as outside 
      the LBP, is needed for a comprehensive understanding of the functionality and 
      interactions of the receptor with these ligands and for development of new small 
      molecules as potential therapeutic drugs.
CI  - Copyright (c) 2011 Elsevier Ltd. All rights reserved.
FAU - Castillo, Hilda S
AU  - Castillo HS
AD  - School of Chemistry & Biochemistry, Parker H. Petit Institute for Bioengineering 
      & Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA. 
      hilda.castillo@chemistry.gatech.edu
FAU - Ousley, Amanda M
AU  - Ousley AM
FAU - Duraj-Thatte, Anna
AU  - Duraj-Thatte A
FAU - Lindstrom, Kelli N
AU  - Lindstrom KN
FAU - Patel, Dina D
AU  - Patel DD
FAU - Bommarius, Andreas S
AU  - Bommarius AS
FAU - Azizi, Bahareh
AU  - Azizi B
LA  - eng
GR  - R01 GM075832/GM/NIGMS NIH HHS/United States
GR  - 1R01GM075832/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20110822
PL  - England
TA  - J Steroid Biochem Mol Biol
JT  - The Journal of steroid biochemistry and molecular biology
JID - 9015483
RN  - 0 (Ligands)
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Recombinant Proteins)
RN  - 1C6V77QF41 (Cholecalciferol)
RN  - 5QU0I8393U (Lithocholic Acid)
RN  - EC 1.13.12.5 (Luciferases, Renilla)
RN  - FXC9231JVH (Calcitriol)
RN  - K848JZ4886 (Cysteine)
SB  - IM
MH  - Binding Sites
MH  - Calcitriol/*chemistry
MH  - Cholecalciferol/*chemistry
MH  - Computer Simulation
MH  - Cysteine/*chemistry
MH  - Genes, Reporter
MH  - HEK293 Cells
MH  - Humans
MH  - Hydrogen Bonding
MH  - Ligands
MH  - Lithocholic Acid/*chemistry
MH  - Luciferases, Renilla/biosynthesis/genetics
MH  - Models, Molecular
MH  - Mutation, Missense
MH  - Protein Binding
MH  - Protein Stability
MH  - Receptors, Calcitriol/*chemistry/genetics/metabolism
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Yeasts
EDAT- 2011/09/03 06:00
MHDA- 2012/02/11 06:00
CRDT- 2011/09/03 06:00
PHST- 2011/04/25 00:00 [received]
PHST- 2011/08/01 00:00 [revised]
PHST- 2011/08/14 00:00 [accepted]
PHST- 2011/09/03 06:00 [entrez]
PHST- 2011/09/03 06:00 [pubmed]
PHST- 2012/02/11 06:00 [medline]
AID - S0960-0760(11)00162-2 [pii]
AID - 10.1016/j.jsbmb.2011.08.003 [doi]
PST - ppublish
SO  - J Steroid Biochem Mol Biol. 2012 Jan;128(1-2):76-86. doi: 
      10.1016/j.jsbmb.2011.08.003. Epub 2011 Aug 22.