9R3H

Structure of liver pyruvate kinase in complex with fluorescent probe 4b

これはPDB形式変換不可エントリーです。

9R3H の概要

• エントリーDOI: 10.2210/pdb9r3h/pdb
• 分子名称: Isoform L-type of Pyruvate kinase PKLR, 1,6-di-O-phosphono-beta-D-fructofuranose, OXALATE ION, ... (7 entities in total)
• 機能のキーワード: pyruvate kinase, fluorescence, allosteric site, transferase
• 由来する生物種: Homo sapiens (human)
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 392037.20

構造登録者

Bogucka, A.,Nilsson, O.,Grotli, M.,Hyvonen, M. (登録日: 2025-05-05, 公開日: 2025-08-13)

主引用文献

Nilsson, O.,Bogucka, A.,Koteles, I.,Haversen, L.,Liljenberg, S.,Rutberg, M.,Hyvonen, M.,Grotli, M.
Fluorescent binding assay for allosteric ligands of liver pyruvate kinase.
Eur.J.Med.Chem., 298:117989-117989, 2025

PubMed Abstract: Environment-sensitive fluorescent probes are indispensable tools for studying biological systems and advancing drug discovery. This study reports the development of 4-sulfamoyl-7-aminobenzoxadiazole (SBD)-based fluorescent probes for the allosteric site of the liver isoform of pyruvate kinase (PKL). By integrating SBD moieties into known activator scaffolds, such as mitapivat and diarylsulfonamide (DASA) ligands, probes for indicator displacement assays were designed to quantify ligand interactions in the allosteric site. Compound 4a displayed dose-dependent fluorescence enhancement in response to PKL binding and was used in a competitive binding assay with unlabelled ligands: mitapivat, TEPP-46, DASA-58 and reported activator 21. Structure-activity relationship (SAR) analysis revealed key structural features influencing activity and fluorescence sensitivity. The probes report selectively on the allosteric site ligands as the binding was not affected by natural ligands, such as ADP, fructose-1,6-bisphosphate (FBP), phosphoenolpyruvate (PEP), and phenylalanine. These findings provide a practical framework for detecting allosteric ligand engagement in PKL and expand the repertoire of molecular tools for advancing PKL-targeted therapies.

PubMed: 40749256
DOI: 10.1016/j.ejmech.2025.117989

実験手法

X-RAY DIFFRACTION (2.1 Å)