5WR1

Covalent bond formation of bifunctional ligand with hPPARg-LBD

Summary for 5WR1

• Entry DOI: 10.2210/pdb5wr1/pdb
• Descriptor: Peroxisome proliferator-activated receptor gamma, 2-[E-(E-16-azido-2-oxidanylidene-hexadec-3-enylidene)amino]ethanoic acid (3 entities in total)
• Functional Keywords: receptor, transcription
• Biological source: Homo sapiens (Human)
• Cellular location: Nucleus: P37231
• Total number of polymer chains: 2
• Total formula weight: 63249.50

Authors

Kojima, H.,Itoh, T.,Yamamoto, K. (deposition date: 2016-11-29, release date: 2017-11-22, Last modification date: 2024-10-16)

Primary citation

Kojima, H.,Itoh, T.,Yamamoto, K.
On-site reaction for PPAR gamma modification using a specific bifunctional ligand
Bioorg. Med. Chem., 25:6492-6500, 2017

PubMed Abstract: Site-specific labeling is an important methodology to elucidate the biological function of a target protein. Here, we report a strategy for site-specific chemical labeling, termed the "on-site reaction". We designed and readily synthesized a bifunctional ligand possessing two reaction sites, an enone and an azide moiety. This strategy involves an on-site conjugate addition reaction with protein followed by a Hüisgen cycloaddition reaction. We demonstrate this strategy by using fluorescein as a probe and peroxisome proliferator activated receptor γ (PPARγ) as a target protein. The reactions were evaluated by ESI-mass analysis and the binding site and modes of binding were revealed by X-ray crystallization analysis. The proposed methodology can easily convert a covalent ligand into chemical tool for protein functional analysis and the identification of drug targets.

PubMed: 29097031
DOI: 10.1016/j.bmc.2017.10.024

Experimental method

X-RAY DIFFRACTION (2.34 Å)