Chemical and Biochemical Reactivity of the Reduced Forms of Nicotinamide Riboside

All life forms require nicotinamide adenine dinucleotide, NAD(+), and its reduced form NADH. They are redox partners in hundreds of cellular enzymatic reactions. Changes in the intracellular levels of total NAD (NAD(+) + NADH) and the (NAD(+)/NADH) ratio can cause cellular dysfunction. When not present in protein complexes, NADH and its phosphorylated form NADPH degrade through intricate mechanisms. Replenishment of a declining total NAD pool can be achieved with biosynthetic precursors that include one of the reduced forms of nicotinamide riboside (NR+), NRH. NRH, like NADH and NADPH, is prone to degradation via oxidation, hydration, and isomerization and, as such, is an excellent model compound to rationalize the nonenzymatic metabolism of NAD(P)H in a biological context. Here, we report on the stability of NRH and its propensity to isomerize and irreversibly degrade. We also report the preparation of two of its naturally occurring isomers, their chemical stability, their reactivity toward NRH-processing enzymes, and their cell-specific cytotoxicity. Furthermore, we identify a mechanism by which NRH degradation causes covalent peptide modifications, a process that could expose a novel type of NADH-protein modifications and correlate NADH accumulation with protein aging. This work highlights the current limitations in detecting NADH's endogenous catabolites and in establishing the capacity for inducing cellular dysfunction.

Automated summary

NAD(+) and NADH are essential for cellular enzymatic reactions and changes in their levels can cause cellular dysfunction. NRH, like NADH and NADPH, is prone to degradation and its degradation can lead to cell-specific cytotoxicity. Further research is needed to understand the mechanisms of NADH degradation and its impact on cell function.