title: ¹⁹F nuclear magnetic resonance screening of glucokinase activators.
abstract: Human hexokinase enzyme IV (EC 2.7.1.1) catalyzes the phosphorylation of glucose and regulates the level of glucose. This enzyme exhibits strong positive cooperativity due to an allosteric transition between an inactive form and a closed active form. This form can be stabilized by activators and, thus, can increase its turnover by a kinetic memory effect characterized by a slow decay to the inactive state. The structural details of this kinetic allostery are known. Several synthetic activators have been reported. We present a preliminary nuclear magnetic resonance (NMR) screening of a chemical library in search of molecules with some affinity for glucokinase (GK). The library, composed of eight molecules with known activity as well as molecules that display no interaction, has been tested using the FAXS (fluorine chemical shift anisotropy and exchange for screening) method, based on monitoring the R2 relaxation of the (19)F spin. To ensure a valid interaction measurement, the enzyme was placed in the presence of glucose and magnesium. The binding signal of one known fluorinated ligand was measured by determining the displacement of the known ligand. This simple measure of the (19)F signal intensity after an 80-ms spin echo correlates nicely with the EC50, opening a route for NMR screening of GK activators.
authors: NMRTEC, 67400 Illkirch Graffenstaden, France.
authors: O
authors: O
authors: Assemat
authors: Division de Pharmacologie Moléculaire et Cellulaire, Institut de Recherches Servier, 78290 Croissy-sur-Seine, France.
authors: M
authors: M
authors: Antoine
authors: PEX Chimie, Institut de recherches SERVIER, 92150 Suresnes, France.
authors: J-M
authors: JM
authors: Fourquez
authors: PEX Chimie, Institut de recherches SERVIER, 92150 Suresnes, France.
authors: M
authors: M
authors: Wierzbicki
authors: PEX Chimie, Institut de recherches SERVIER, 92150 Suresnes, France.
authors: Y
authors: Y
authors: Charton
authors: PEX Chimie, Institut de recherches SERVIER, 92150 Suresnes, France.
authors: P
authors: P
authors: Hennig
authors: PEX Chimie, Institut de recherches SERVIER, 92150 Suresnes, France.
authors: F
authors: F
authors: Perron-Sierra
authors: Division de Pharmacologie Moléculaire et Cellulaire, Institut de Recherches Servier, 78290 Croissy-sur-Seine, France.
authors: G
authors: G
authors: Ferry
authors: Division de Pharmacologie Moléculaire et Cellulaire, Institut de Recherches Servier, 78290 Croissy-sur-Seine, France. Electronic address: jean.boutin@fr.netgrs.com.
authors: J A
authors: JA
authors: Boutin
authors: NMRTEC, 67400 Illkirch Graffenstaden, France; Institut de génétique et de biologie moléculaire et cellulaire, 67404 Illkirch Cedex, France.
authors: M-A
authors: MA
authors: Delsuc
mesh_terms: D004353:Drug Evaluation, Preclinical; D020536:Enzyme Activators / Q000494:pharmacology*; D005941:Glucokinase / Q000378:metabolism*; D054879:Halogenation; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*
chemical_list: D020536:Enzyme Activators; D008024:Ligands; D005941:Glucokinase
keywords: (19)F spin signal; Enzyme-activating assay; Glucokinase; Ligands; NMR