Computational and experimental studies were performed to determine the role of N(3)H hydrogen bonding in controlling flavin redox chemistry. B3LYP calculations of a lumiflavin-DMF complex indicate that hydrogen bonding to N(3)H is redox state dependent: hydrogen bonds to this position are formed in the oxidized state, and released in the flavin radical anion system due to increased electron density in the reduced species. Electrochemical studies of flavin and N(3)-methyl flavin in hydrogen bonding and non-hydrogen bonding solvents confirm this prediction, demonstrating that N(3)H hydrogen bonding modulates the potential of flavin reduction to the corresponding radical anion by 80 mV (1.8 kcal/mol). Variable-temperature electrochemical studies were also performed to establish the enthalpic and entropic effects of N(3)H hydrogen bonding on flavin reduction.