6JEZ

Covalent labeling of rVDR-LBD by turn-on fluorescent probe mediated by conjugate addition and cyclization

6JEZ の概要

• エントリーDOI: 10.2210/pdb6jez/pdb
• 分子名称: Vitamin D3 receptor,Vitamin D3 receptor, Mediator of RNA polymerase II transcription subunit 1, (1R,3R)-5-(2-((1R,3aS,7aR,E)-1-((R)-6-hydroxy-6-methylheptan-2-yl)-7a-methyloctahydro-4H-inden-4-ylidene)ethylidene)-2- methylenecyclohexane-1,3-diol, ... (5 entities in total)
• 機能のキーワード: hormone, turn-on fluorescent probe, tcc probe, covalent labeling, histidine
• 由来する生物種: Rattus norvegicus (Rat); 詳細
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 32799.83

構造登録者

Kojima, H.,Yamamoto, K.,Itoh, T. (登録日: 2019-02-07, 公開日: 2020-02-12, 最終更新日: 2023-11-22)

主引用文献

Kojima, H.,Fujita, Y.,Takeuchi, R.,Ikebe, Y.,Ohashi, N.,Yamamoto, K.,Itoh, T.
Cyclization Reaction-Based Turn-on Probe for Covalent Labeling of Target Proteins.
Cell Chem Biol, 27:334-, 2020

PubMed Abstract: Fluorescent molecules have contributed to basic biological research but there are currently only a limited number of probes available for the detection of non-enzymatic proteins. Here, we report turn-on fluorescent probes mediated by conjugate addition and cyclization (TCC probes). These probes react with multiple amino acids and exhibit a 36-fold greater emission intensity after reaction. We analyzed the reactions between TCC probes and nuclear receptors by electrospray ionization mass spectrometry, X-ray crystallography, spectrofluorometry, and fluorescence microscopy. In vitro analysis showed that probes consisting of a protein ligand and TCC could label vitamin D receptor and peroxisome proliferator-activated receptor γ. Moreover, we demonstrated that not only a ligand unit but also a peptide unit can label the target protein in a complex mixture.

PubMed: 31991094
DOI: 10.1016/j.chembiol.2020.01.006

実験手法

X-RAY DIFFRACTION (2.3 Å)