Mechanism of O2 activation and substrate hydroxylation in noncoupled binuclear copper monooxygenases.

ABSTRACT: Peptidylglycine ?-hydroxylating monooxygenase (PHM) and dopamine ?-monooxygenase (D?M) are copper-dependent enzymes that are vital for neurotransmitter regulation and hormone biosynthesis. These enzymes feature a unique active site consisting of two spatially separated (by 11 Å in PHM) and magnetically noncoupled copper centers that enables 1e- activation of O2 for hydrogen atom abstraction (HAA) of substrate C-H bonds and subsequent hydroxylation. Although the structures of the resting enzymes are known, details of the hydroxylation mechanism and timing of long-range electron transfer (ET) are not clear. This study presents density-functional calculations of the full reaction coordinate, which demonstrate: (i) the importance of the end-on coordination of superoxide to Cu for HAA along the triplet spin surface; (ii) substrate radical rebound to a CuII hydroperoxide favors the proximal, nonprotonated oxygen; and (iii) long-range ET can only occur at a late step with a large driving force, which serves to inhibit deleterious Fenton chemistry. The large inner-sphere reorganization energy at the ET site is used as a control mechanism to arrest premature ET and dictate the correct timing of ET.

SUBMITTER: Cowley RE 

PROVIDER: S-EPMC5087064 | biostudies-literature | 2016 Oct

REPOSITORIES: biostudies-literature