Toward Quantitative Measurements of Enzyme Kinetics by Dissolution Dynamic Nuclear Polarization.
Dissolution dynamic nuclear polarization (D-DNP) experiments enabled us to study the kinetics of the enzymatic phosphorylation reaction of glucose to form glucose-6-phosphate (G6P) by hexokinase (HK), with or without the presence of an excess of G6P, which is known to be an inhibitor of the enzyme. Against all expectations, our observations demonstrate that the phosphorylation of both α and β glucose anomers occurs with comparable kinetics. The catalytic constant of the reaction was estimated based on a simple kinetic model tailored for hyperpolarized systems.
†École Normale Supérieure-PSL Research University, Département de Chimie, 24 rue Lhomond, F-75005 Paris, France.|‡Sorbonne Universités, UPMC Univ Paris 06, LBM, 4 place Jussieu, F-75005, Paris, France.|§CNRS, UMR 7203 LBM, F-75005, Paris, France.
Emeric
E
Miclet
†École Normale Supérieure-PSL Research University, Département de Chimie, 24 rue Lhomond, F-75005 Paris, France.|‡Sorbonne Universités, UPMC Univ Paris 06, LBM, 4 place Jussieu, F-75005, Paris, France.|§CNRS, UMR 7203 LBM, F-75005, Paris, France.
Daniel
D
Abergel
∥Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne (EPFL), Batochime, CH-1015 Lausanne, Switzerland.
Aurélien
A
Bornet
∥Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne (EPFL), Batochime, CH-1015 Lausanne, Switzerland.
Jonas
J
Milani
†École Normale Supérieure-PSL Research University, Département de Chimie, 24 rue Lhomond, F-75005 Paris, France.|‡Sorbonne Universités, UPMC Univ Paris 06, LBM, 4 place Jussieu, F-75005, Paris, France.|§CNRS, UMR 7203 LBM, F-75005, Paris, France.|∥Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne (EPFL), Batochime, CH-1015 Lausanne, Switzerland.|⊥Bruker BioSpin AG, Industriestrasse 26, 8117 Fällanden, Switzerland.
Sami
S
Jannin
†École Normale Supérieure-PSL Research University, Département de Chimie, 24 rue Lhomond, F-75005 Paris, France.|‡Sorbonne Universités, UPMC Univ Paris 06, LBM, 4 place Jussieu, F-75005, Paris, France.|§CNRS, UMR 7203 LBM, F-75005, Paris, France.|∥Institut des Sciences et Ingénierie Chimiques, Ecole Polytechnique Fédérale de Lausanne (EPFL), Batochime, CH-1015 Lausanne, Switzerland.
Geoffrey
G
Bodenhausen
dissolution dynamic nuclear polarization; enzyme kinetics; glucose-6-phosphate; hexokinase; hyperpolarization; nuclear magnetic resonance