PMID- 9153406
OWN - NLM
STAT- MEDLINE
DCOM- 19970609
LR  - 20131121
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 36
IP  - 19
DP  - 1997 May 13
TI  - The isolation and characterization of purified heterocomplexes of recombinant 
      retinoic acid receptor and retinoid X receptor ligand binding domains.
PG  - 5669-76
AB  - Retinoic acid exerts many of its biological effects by interaction with 
      heterocomplexes of nuclear retinoic acid receptors (RARs) and retinoid X 
      receptors (RXRs). To further examine this interaction, a glutathione 
      S-transferase (GST) fusion protein containing the ligand binding domain of human 
      RXR alpha has been used to copurify the ligand binding domain of human RAR gamma 
      by affinity chromatography over glutathione-agarose. Complexes of recombinant 
      RAR-RXR ligand binding domains retaining full ligand binding capacity were 
      purified, and their interactions with various retinoids were characterized by 
      fluorometric titration and photoaffinity labeling. Analyses of the distribution 
      of limiting amounts of [3H]-all-trans-retinoic acid between cytoplasmic retinoic 
      acid binding proteins, CRABP-I and CRABP-II, and the purified heterocomplexes 
      indicate that all-trans-retinoic acid binds with comparable affinity to CRABP-I 
      and the heterocomplexes, but with approximately 10-fold less affinity to 
      CRABP-II. The aromatic retinoid acitretin, which is used in the treatment of 
      psoriasis, binds relatively poorly to the purified heterocomplexes, although it 
      binds with high affinity to the CRABPs. Acitretin displaces 
      [3H]-all-trans-retinoic acid from the CRABPs and increases retinoic acid 
      occupancy of the heterocomplexes. These results suggest that certain retinoids 
      could potentially perturb the distribution of endogenous retinoic acid between 
      the CRABPs and the nuclear receptors and thus affect retinoid signaling. The 
      purified recombinant complexes should provide a useful model system for further 
      structural analysis of the dimerization interface between the RAR and RXR ligand 
      binding domains.
FAU - Tian, K
AU  - Tian K
AD  - Department of Medicine, Washington University School of Medicine, St. Louis, 
      Missouri 63110, USA.
FAU - Norris, A W
AU  - Norris AW
FAU - Lin, C L
AU  - Lin CL
FAU - Li, E
AU  - Li E
LA  - eng
GR  - 5-T32GM07200/GM/NIGMS NIH HHS/United States
GR  - DK40172/DK/NIDDK NIH HHS/United States
GR  - DK49684/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Affinity Labels)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Ligands)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Receptors, Retinoic Acid)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Retinoid X Receptors)
RN  - 0 (Retinoids)
RN  - 0 (Transcription Factors)
RN  - 0 (retinoic acid binding protein I, cellular)
RN  - 0 (retinoic acid binding protein II, cellular)
RN  - 5688UTC01R (Tretinoin)
RN  - LCH760E9T7 (Acitretin)
SB  - IM
MH  - Acitretin/pharmacology
MH  - Affinity Labels
MH  - Animals
MH  - DNA-Binding Proteins/*chemistry/*isolation & purification/metabolism
MH  - Ligands
MH  - Macromolecular Substances
MH  - Mice
MH  - Protein Structure, Tertiary
MH  - Receptors, Retinoic Acid/*chemistry/drug effects/*isolation & 
      purification/metabolism
MH  - Recombinant Fusion Proteins/*chemistry/*isolation & purification/metabolism
MH  - Retinoid X Receptors
MH  - Retinoids/chemistry/metabolism
MH  - Spectrometry, Fluorescence
MH  - Titrimetry
MH  - Transcription Factors/*chemistry/*isolation & purification/metabolism
MH  - Tretinoin/chemistry/metabolism
EDAT- 1997/05/13 00:00
MHDA- 1997/05/13 00:01
CRDT- 1997/05/13 00:00
PHST- 1997/05/13 00:00 [pubmed]
PHST- 1997/05/13 00:01 [medline]
PHST- 1997/05/13 00:00 [entrez]
AID - bi9627020 [pii]
AID - 10.1021/bi9627020 [doi]
PST - ppublish
SO  - Biochemistry. 1997 May 13;36(19):5669-76. doi: 10.1021/bi9627020.