TY - JOUR
T1 - HX531, a retinoid X receptor antagonist, inhibited the 9-cis retinoic acid-induced binding with steroid receptor coactivator-1 as detected by surface plasmon resonance
AU - Kanayasu-Toyoda, Toshie
AU - Fujino, Tomofumi
AU - Oshizawa, Tadashi
AU - Suzuki, Takayoshi
AU - Nishimaki-Mogami, Tomoko
AU - Sato, Yoji
AU - Sawada, Jun Ichi
AU - Inoue, Kazuhide
AU - Shudo, Koichi
AU - Ohno, Yasuo
AU - Yamaguchi, Teruhide
N1 - Funding Information:
This work was supported by a grant (MF-16) from the Organization for Pharmaceutical Safety and Research and a grant from the Japan Health Sciences Foundation.
Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2005/3
Y1 - 2005/3
N2 - HX531 is a retinoid X receptor (RXR) antagonist that inhibits 9-cis retinoic acid-induced neutrophilic differentiation of HL-60 cells. In order to elucidate the inhibitory mechanism of HX531, we have developed a novel ligand sensor assay for RXR in which the receptor-coactivator interaction is directly monitored using surface plasmon resonance (SPR) biosensor technology. A 20-mer peptide from steroid receptor coactivator-1 (SRC-1), containing nuclear receptor interaction motif LXXLL was immobilized on the surface of a BIAcore sensor chip. Injection of human recombinant RXR with or without 9-cis retinoic acid resulted in ligand-dependent interaction with the SRC-1 peptide. Kinetic analysis revealed dissociation constants (KD) of 9-cis RA-preincubated RXR to SRC-1 was 5.92 × 10-8 M. Using this technique, we found that 1 μM HX531 reduced the ka value of liganded-RXR with SRC-1, suggesting that HX531 reduced the affinity of RXR to SRC-1. This SPR assay system was applied to obtain quantitative kinetic data of RXR ligand binding to the SRC-1 peptide and the alteration of these data by antagonists.
AB - HX531 is a retinoid X receptor (RXR) antagonist that inhibits 9-cis retinoic acid-induced neutrophilic differentiation of HL-60 cells. In order to elucidate the inhibitory mechanism of HX531, we have developed a novel ligand sensor assay for RXR in which the receptor-coactivator interaction is directly monitored using surface plasmon resonance (SPR) biosensor technology. A 20-mer peptide from steroid receptor coactivator-1 (SRC-1), containing nuclear receptor interaction motif LXXLL was immobilized on the surface of a BIAcore sensor chip. Injection of human recombinant RXR with or without 9-cis retinoic acid resulted in ligand-dependent interaction with the SRC-1 peptide. Kinetic analysis revealed dissociation constants (KD) of 9-cis RA-preincubated RXR to SRC-1 was 5.92 × 10-8 M. Using this technique, we found that 1 μM HX531 reduced the ka value of liganded-RXR with SRC-1, suggesting that HX531 reduced the affinity of RXR to SRC-1. This SPR assay system was applied to obtain quantitative kinetic data of RXR ligand binding to the SRC-1 peptide and the alteration of these data by antagonists.
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U2 - 10.1016/j.jsbmb.2004.11.007
DO - 10.1016/j.jsbmb.2004.11.007
M3 - Article
C2 - 15857749
AN - SCOPUS:20944439398
SN - 0960-0760
VL - 94
SP - 303
EP - 309
JO - Journal of Steroid Biochemistry
JF - Journal of Steroid Biochemistry
IS - 4
ER -
