Recombinant human retinoic acid receptor beta. Binding of synthetic retinoids and transcriptional activation.
All-trans-retinoic acid mediates cell growth and differentiation by binding to and then activating nuclear retinoid receptor proteins that regulate gene transcription. Recombinant human retinoic acid receptor beta was cloned and expressed in Escherichia coli as a fusion protein rMBP-RAR beta with maltose-binding protein to facilitate purification. After isolation from bacterial lysates, rMBP-RAR beta was used for binding with selected retinoids. Scatchard analysis with [11,12-3H2]all-trans-retinoic acid gave a Kd of 0.34 nM. Competitive binding studies with a series of conformationally restricted aromatic retinoids indicated that the Ki values for binding to rMBP-RAR beta correlated with the logs of the EC50 values for gene transcriptional activation (p < or = 0.05) and with those for the relative activation compared to that of all-trans-retinoic acid (p < or = 0.01). Inspection of binding-activation correlation diagrams indicates candidate structures for improved retinoid agonists or antagonists.
Lombardo
A
A
Cancer Research Center, La Jolla Cancer Research Foundation, California 92037.
Costa
E
E
Chao
W R
WR
Toll
L
L
Hobbs
P D
PD
Jong
L
L
Lee
M O
MO
Pfahl
M
M
Ely
K R
KR
Dawson
M I
MI
D000595:Amino Acid Sequence; D001667:Binding, Competitive; D003001:Cloning, Molecular; D004926:Escherichia coli; D006801:Humans; D008969:Molecular Sequence Data; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012176:Retinoids / Q000378:metabolism*; D015533:Transcriptional Activation
D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D012176:Retinoids; C082925:retinoic acid receptor beta