6JF0

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

Summary for 6JF0

• Entry DOI: 10.2210/pdb6jf0/pdb
• Descriptor: Peroxisome proliferator-activated receptor gamma, methyl (2~{S})-3-[4-[3-(4-methoxy-2-oxidanyl-phenyl)prop-2-ynoyloxy]phenyl]-2-[[2-(phenylcarbonyl)phenyl]amino]propanoate, 7-methoxychromen-2-one, ... (4 entities in total)
• Functional Keywords: receptor, covalent labeling, cysteine, turn-on fluorescent probe, tcc probe, nuclear protein
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 2
• Total formula weight: 63624.78

Authors

Kojima, H.,Yamamoto, K.,Itoh, T. (deposition date: 2019-02-07, release date: 2020-02-12, Last modification date: 2023-11-22)

Primary citation

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

Experimental method

X-RAY DIFFRACTION (3.4 Å)