3CTB

Tethered PXR-LBD/SRC-1p apoprotein

3CTB の概要

• エントリーDOI: 10.2210/pdb3ctb/pdb
• 関連するPDBエントリー: 1ILG 1ILH 1NRL 3CTC
• 分子名称: Pregnane X receptor, Linker, Steroid receptor coactivator 1 (2 entities in total)
• 機能のキーワード: pxr, src-1, tethered, engineered, drug-drug interactions, dna-binding, metal-binding, nucleus, receptor, transcription regulation, zinc-finger, transcription, transferase
• 由来する生物種: Homo sapiens (human); 詳細
• 細胞内の位置: Nucleus : Q15788
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 78368.07

構造登録者

Lesburg, C.A. (登録日: 2008-04-11, 公開日: 2008-12-02, 最終更新日: 2023-08-30)

主引用文献

Wang, W.,Prosise, W.W.,Chen, J.,Taremi, S.S.,Le, H.V.,Madison, V.,Cui, X.,Thomas, A.,Cheng, K.C.,Lesburg, C.A.
Construction and characterization of a fully active PXR/SRC-1 tethered protein with increased stability
Protein Eng.Des.Sel., 21:425-433, 2008

PubMed Abstract: The nuclear xenobiotic receptor PXR is a ligand-inducible transcription factor regulating drug-metabolizing enzymes and transporters and a master switch mediating potentially adverse drug-drug interactions. In addition to binding a coactivator protein such as SRC-1, the C-terminal ligand-binding domain (LBD) is solely responsible for ligand recognition and thus the ligand-dependent downstream effects. In an effort to facilitate structural studies of PXR to understand and abolish the interactions between PXR and its ligands, several recombinant PXR/SRC-1 constructs were designed and evaluated for expression, stability and activity. Expression strategies employing either dual expression or translationally coupled bicistronic expression were found to be unsuitable for producing stable PXR in a stochiometric complex with a peptide derived from SRC-1 (SRC-1p). A single polypeptide chain encompassing PXR and SRC-1p tethered with a peptidyl linker was designed to promote intramolecular complex formation. This tethered protein was overexpressed as a soluble protein and required no additional SRC-1p for further stabilization. X-ray crystal structures in the presence and absence of the known PXR agonist SR-12813 were determined to high resolution. In addition, a circular dichroism-based binding assay was developed to allow rapid evaluation of PXR ligand affinity, making this tethered protein a convenient and effective reagent for the rational attenuation of drug-induced PXR-mediated metabolism.

PubMed: 18456871
DOI: 10.1093/protein/gzn017

実験手法

X-RAY DIFFRACTION (2 Å)