Fry et al.
oxidase13-15 are also asymmetric. More recently, stoichio-
metric CO binding studies and IR spectroscopic measure-
ments helped to characterize the structural and functional
inequivalence of the two copper(I) centers (CuA t CuH, CuB
t CuM) of reduced dopamine-â-monoxygenase (DâH)16-18
and peptidylglycine monooxygenase (PHM).19-22 In addition,
CO facilitated the characterization/description of an unprec-
edented S-donor methionine ligand at the CuB/CuM site and
aided in the elucidation of the ligand environment for the
CuI versus CuII centers.
Chart 1
Although CO has been most commonly utilized to
determine active site structure in copper enzymes, it has also
helped in elucidating mechanistic pathways. For example,
the ability of CO to bind to both PHM metal sites,
surprisingly even CuH when peptidylglycine substrates bind,22
at the time led to new hypotheses for probing the enzyme
mechanism and dynamics (i.e., superoxide channeling20,22 vs
hydrogen tunneling for the substrate hydrogen atom abstrac-
tion23-27) of the uncoupled metal centers. In addition, the
course of CO and presumably O2 reactivity in the hemea3/
CuB active site of cytochrome c oxidase (CcO) has been
probed through laser flash photolysis (LFP).28-33 Upon the
photorelease of carbon monoxide from hemea3, CO rapidly
(ps) moves to CuIB and subsequently escapes or returns to
the hemea3 on a millisecond time-scale, suggesting that CuIB
can act as a doorway for substrate O2-binding to hemea3 in
this binuclear active site of CcO. LFP is a common
methodology used to probe heme protein-actives-site envi-
ronments giving kinetic and thermodynamic insights.33-35
Whereas CO photoejection has yet to be observed from a
protein that has only copper as the active-site metal, we
recently communicated that metal-to-ligand charge-transfer
(MLCT) excitation of at least one type of copper(I) carbonyl
complex, [CuI(H-tmpa)(CO)]+ (1H-CO; TMPA ) tris(2-
pyridylmethyl)amine); Chart 1), resulted in the efficient
release of CO.36 The CO rebinding was quantitative, and no
net photochemistry was observed. The rate constant for CO
rebinding following flash photolysis of 1H-CO in tetrahy-
drofuran solvent (THF) was determined to approach a
diffusion-controlled value, kCO ) 1.9 × 109 M-1 s-1 at
25 °C. We were also successful at applying the flash-and-
trap technique, pioneered by Gibson and co-workers for heme
protein analysis.37 Light excitation (λex ) 355 nm) of 1H-
CO in the presence of O2 in THF resulted in the transient
appearance of a primary copper(I)-dioxygen adduct, a
cupric-superoxo (CuII(O2•-)) species.38 The temperature
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242 Inorganic Chemistry, Vol. 47, No. 1, 2008