Quantum chemical calculations of the NHA bound nitric oxide synthase active site: O₂ binding and implications for the catalytic mechanism

Density functional theory has been employed to model the binding of the intermediate substrate NHA, by nitric oxide synthases, In particular, the orientation and interactions of possibly catalytically important substrate hydrogens, with and without molecular oxygen bound to the active site heme group, are considered. Without O₂, three possible conformers have been found, with the energetically most favored structure being that in which both protons of the -NHOH moiety of NHA are directed toward the heme group. With oxygen bound, four different structures were found, The energetically lowest structure is again found to have both hydrogens of the -NHOH group pointing toward the heme group, thus forming hydrogen bonds between -NH- and the terminal oxygen, and between -OH and the inner oxygen of the heme-O₂ group. In addition, unprotonated structures of the substrate bound to the active site are considered and the proton affinity calculated.