Sasaki T, Morisaki N, Iwasaki S, Kagechika H, Fukasawa H, Shudo K, Shida Y, Hashimoto Y
Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.
Biol Pharm Bull. 1997 Aug;20(8):913-6. doi: 10.1248/bpb.20.913.
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.
设计并合成了两种用于视黄酸受体（RAR）α的光亲和标记探针，即4-[(3-(3-(三氟甲基)-3H-重氮丙啶-3-基)苯基)羧酰胺基]苯甲酸（3DIAM）及其对位异构体（4DIAM）。这两种化合物对重组RARα（MBP-RARα/E）都具有高亲和力，并与其同源配体结合位点共价结合。通过内蛋白酶组合方法确定，3DIAM标记的MBP-RARα/E位点位于RARα配体结合域的螺旋11中，根据视黄酸/RARγ复合物的报道晶体结构，该位置被认为是配体结合的位置。
