Direct coordination of pterin to FeII enables neurotransmitter biosynthesis in the pterin-dependent hydroxylases

The pterin-dependent nonheme iron enzymes hydroxylate aromatic amino acids to perform the biosynthesis of neurotransmitters to maintain proper brain function. These enzymes activate oxygen using a pterin cofactor and an aromatic amino acid substrate bound to the FeII active site to form a highly reactive Fe-IV = O species that initiates substrate oxidation. In this study, using tryptophan hydroxylase, we have kinetically generated a pre-Fe-IV = O intermediate and characterized its structure as a Fe-II-peroxy-pterin species using absorption, M?ssbauer, resonance Raman, and nuclear resonance vibrational spectroscopies. From parallel characterization of the pterin cofactor and tryptophan substrate?bound ternary Fe-II active site before the O-2 reaction (including magnetic circular dichroism spectroscopy), these studies both experimentally define the mechanism of Fe-IV = O formation and demonstrate that the carbonyl functional group on the pterin is directly coordinated to the Fe-II site in both the ternary complex and the peroxo intermediate. Reaction coordinate calculations predict a 14 kcal/mol reduction in the oxygen activation barrier due to the direct binding of the pterin carbonyl to the Fe-II site, as this interaction provides an orbital pathway for efficient electron transfer from the pterin cofactor to the iron center. This direct coordination of the pterin cofactor enables the biological function of the pterin-dependent hydroxylases and demonstrates a unified mechanism for oxygen activation by the cofactor-dependent nonheme iron enzymes.

Automated summary

The study identified a pre-Fe-IV = O intermediate in the reaction with tryptophan hydroxylase, characterized as a Fe-II-peroxy-pterin species. It was revealed that direct coordination of the pterin carbonyl to the Fe-II site may reduce the oxygen activation barrier according to reaction coordinate calculations.