Correlation of temperature dependence of hydride kinetic isotope effects with donor-acceptor distances in two solvents of different polarities

Recently observed nearly temperature ( T )-independent kinetic isotope effects (KIEs) in wild-type enzymes and T -dependent KIEs in variants were used to suggest that H-tunneling in enzymes is assisted by the fast protein vibrations that help sample short donor-acceptor distances (DADs). This supports the recently proposed role of protein vibrations in DAD sampling catalysis. However, use of T -dependence of KIEs to suggest DAD sampling associated with protein vibrations is debated. We have formulated a hypothesis regarding the correlation and designed experiments in solution to investigate it. The hypothesis is, a more rigid system with shorter DAD TRS 's at the tunneling ready states (TRSs) gives rise to a weaker T -dependence of KIEs, i.e. , a smaller Δ E a (= E aD − E aH ). In a former work, the solvent effects of acetonitrile versus chloroform on the Δ E a of NADH/NAD + model reactions were determined, and the DAD PRC 's of the productive reactant complexes (PRCs) were computed to substitute the DAD TRS for the DAD TRS -Δ E a correlation study. A smaller Δ E a was found in the more polar acetonitrile where the positively charged PRC is better solvated and has a shorter DAD PRC , indirectly supporting the hypothesis. In this work, the TRS structures of different DAD TRS 's for the hydride tunneling reaction from 1,3-dimethyl-2-phenylimidazoline to 10-methylacridinium were computed. The N-CH 3 /CD 3 secondary KIEs on both reactants were calculated and fitted to the observed values to find the DAD TRS order in both solutions. It was found that the equilibrium DAD TRS is shorter in acetonitrile than in chloroform. Results directly support the DAD TRS -Δ E a correlation hypothesis as well as the explanation that links T -dependence of KIEs to DAD sampling catalysis in enzymes. Shorter donor-acceptor distances (DADs) in hydride transfer reactions give rise to a weaker temperature ( T ) dependence of KIEs. This supports the debated proposal that links T -dependence of KIEs to DAD sampling catalysis studies in enzymes.