title: Photoaffinity labeling of the ligand-interacting helix of the retinoic acid receptor alpha.
abstract: 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.
authors.affiliation: Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Sasaki
authors.forename: N
authors.initials: N
authors.lastname: Morisaki
authors.forename: S
authors.initials: S
authors.lastname: Iwasaki
authors.forename: H
authors.initials: H
authors.lastname: Kagechika
authors.forename: H
authors.initials: H
authors.lastname: Fukasawa
authors.forename: K
authors.initials: K
authors.lastname: Shudo
authors.forename: Y
authors.initials: Y
authors.lastname: Shida
authors.forename: Y
authors.initials: Y
authors.lastname: Hashimoto
mesh_terms: 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
chemical_list: 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