PMID- 8267583
OWN - NLM
STAT- MEDLINE
DCOM- 19940125
LR  - 20190612
IS  - 0006-291X (Print)
IS  - 0006-291X (Linking)
VI  - 197
IP  - 2
DP  - 1993 Dec 15
TI  - Purified human vitamin D receptor overexpressed in E. coli and baculovirus 
      systems does not bind 1,25-dihydroxyvitamin D3 hormone efficiently unless 
      supplemented with a rat liver nuclear extract.
PG  - 478-85
AB  - We report here that highly purified human vitamin D receptor (hVDR) derived from 
      E. coli or baculovirus expression systems does not exhibit saturable, high 
      affinity 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) ligand binding when these 
      preparations alone are analyzed. Inclusion of rat liver nuclear extract, which 
      does not itself contain detectable 1,25(OH)2D3 binding activity, is required to 
      endow hVDR isolated from bacterial or insect cells with the property of high 
      affinity hormone binding (Kd = 0.13-0.22 nM). This observation should facilitate 
      the valid assay of 1,25(OH)2D3 binding activity and kinetics in samples of 
      overexpressed hVDR. Moreover, since rat liver nuclear extract contains retinoid X 
      receptors and possibly other auxiliary factors capable of forming heterodimers 
      with hVDR that in turn associate with vitamin D responsive elements, we 
      hypothesize that like DNA binding, 1,25(OH)2D3 binding to hVDR requires the 
      cooperation of a co-receptor or some uncharacterized receptor 
      activating/stabilizing factor.
FAU - Nakajima, S
AU  - Nakajima S
AD  - Department of Biochemistry, College of Medicine, University of Arizona, Tucson 
      85724.
FAU - Hsieh, J C
AU  - Hsieh JC
FAU - MacDonald, P N
AU  - MacDonald PN
FAU - Haussler, C A
AU  - Haussler CA
FAU - Galligan, M A
AU  - Galligan MA
FAU - Jurutka, P W
AU  - Jurutka PW
FAU - Haussler, M R
AU  - Haussler MR
LA  - eng
GR  - AR-15781/AR/NIAMS NIH HHS/United States
GR  - DK-33351/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochem Biophys Res Commun
JT  - Biochemical and biophysical research communications
JID - 0372516
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Receptors, Calcitriol)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tissue Extracts)
RN  - 104982-03-8 (Osteocalcin)
RN  - FXC9231JVH (Calcitriol)
SB  - IM
MH  - Animals
MH  - Baculoviridae/genetics
MH  - Calcitriol/*metabolism
MH  - Cell Line
MH  - Cell Nucleus/*metabolism
MH  - Cloning, Molecular
MH  - DNA-Binding Proteins/metabolism
MH  - Escherichia coli/metabolism
MH  - Humans
MH  - Kinetics
MH  - Liver/*metabolism
MH  - Moths
MH  - Osteocalcin/genetics
MH  - Rats
MH  - Receptors, Calcitriol/biosynthesis/isolation & purification/*metabolism
MH  - Recombinant Proteins/biosynthesis/isolation & purification/metabolism
MH  - Tissue Extracts/*metabolism
MH  - Transfection
EDAT- 1993/12/15 00:00
MHDA- 1993/12/15 00:01
CRDT- 1993/12/15 00:00
PHST- 1993/12/15 00:00 [pubmed]
PHST- 1993/12/15 00:01 [medline]
PHST- 1993/12/15 00:00 [entrez]
AID - S0006291X83725040 [pii]
AID - 10.1006/bbrc.1993.2504 [doi]
PST - ppublish
SO  - Biochem Biophys Res Commun. 1993 Dec 15;197(2):478-85. doi: 
      10.1006/bbrc.1993.2504.