Triple Isotope Effects Support Concerted Hydride and Proton Transfer and Promoting Vibrations in Human Heart Lactate Dehydrogenase.

ABSTRACT: Transition path sampling simulations have proposed that human heart lactate dehydrogenase (LDH) employs protein promoting vibrations (PPVs) on the femtosecond (fs) to picosecond (ps) time scale to promote crossing of the chemical barrier. This chemical barrier involves both hydride and proton transfers to pyruvate to form l-lactate, using reduced nicotinamide adenine dinucleotide (NADH) as the cofactor. Here we report experimental evidence from three types of isotope effect experiments that support coupling of the promoting vibrations to barrier crossing and the coincidence of hydride and proton transfer. We prepared the native (light) LDH and a heavy LDH labeled with ¹³C, ¹⁵N, and nonexchangeable ²H (D) to perturb the predicted PPVs. Heavy LDH has slowed c

SUBMITTER: Wang Z 

PROVIDER: S-EPMC5244818 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature