Photoaffinity labeling of the ligand-interacting helix of the retinoic acid receptor alpha.
Two photoaffinity-labeling probes for retinoic acid receptor (RAR) alpha, 4-[(3-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)carboxamido]ben zoic acid (3DIAM) and its para-isomer (4DIAM), were designed and synthesized. Both compounds possess high affinity for recombinant RAR alpha (MBP-RAR alpha/E) and bind covalently to its cognate ligand-binding site. The labeled site of MBP-RAR alpha/E with 3DIAM was determined, by the endoproteinase combination method, to be located in helix 11 of the ligand-binding domain of RAR alpha, which is the position at which the ligand is considered to bind, on the basis of the reported crystal structure of the retinoic acid/RAR gamma complex.
Sasaki
T
T
Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.
Morisaki
N
N
Iwasaki
S
S
Kagechika
H
H
Fukasawa
H
H
Shudo
K
K
Shida
Y
Y
Hashimoto
Y
Y
D001389:Azirines / Q000138:chemical synthesis; D001549:Benzamides / Q000138:chemical synthesis; D001665:Binding Sites; D010450:Endopeptidases / Q000737:chemistry; D008024:Ligands; D019920:Photoaffinity Labels; D018168:Receptors, Retinoic Acid / Q000737:chemistry*; D011994:Recombinant Proteins / Q000378:metabolism
C108791:4-((3-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)carboxamido)benzoic acid; C108792:4-((4-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenyl)carboxamido)benzoic acid; D001389:Azirines; D001549:Benzamides; D008024:Ligands; D019920:Photoaffinity Labels; D018168:Receptors, Retinoic Acid; D011994:Recombinant Proteins; D010450:Endopeptidases