Abstract
Binuclear non-heme iron enzymes activate O2 to perform diverse chemistries. Three different structural mechanisms of O2 binding to a coupled binuclear iron site have been identified utilizing variable-temperature, variable-field magnetic circular dichroism spectroscopy (VTVH MCD). For the μ-OH-bridged Fe(II)2 site in hemerythrin, O2 binds terminally to a five-coordinate Fe(II) center as hydroperoxide with the proton deriving from the μ-OH bridge and the second electron transferring through the resulting μ-oxo superexchange pathway from the second coordinatively saturated Fe(II) center in a proton-coupled electron transfer process. For carboxylate-only-bridged Fe(II)2 sites, O2 binding as a bridged peroxide requires both Fe(II) centers to be coordinatively unsaturated and has good frontier orbital overlap with the two orthogonal O2 π* orbitals to form peroxo-bridged Fe(III)2 intermediates. Alternatively, carboxylate-only-bridged Fe(II)2 sites with only a single open coordination position on an Fe(II) enable the one-electron formation of Fe(III)–O2− or Fe(III)–NO− species. Finally, for the peroxo-bridged Fe(III)2 intermediates, further activation is necessary for their reactivities in one-electron reduction and electrophilic aromatic substitution, and a strategy consistent with existing spectral data is discussed.
Keywords: Binuclear non-heme iron enzymes, O2 activation, Variable-temperature, variable-field magnetic circular dichroism, Frontier molecular orbitals, Peroxide activation
Introduction
The binuclear non-heme iron proteins and enzymes form expanding classes [1, 2] that play key roles in antibiotic biosynthesis [3, 4] are targets for drug inhibition [5, 6] and have important roles in biotechnology [7, 8] and bioremediation [9]. On a molecular level, they reversibly bind dioxygen [10], activate it for H-atom abstraction [11], desaturation [12], hydroxylation [13–15], and electrophilic aromatic substitution (EAS) [16, 17], and reduce O2 to H2O and NO to N2O [18]. In contrast to the mononuclear non-heme iron enzymes which often require a cofactor [19, 20], the second Fe of the binuclear non-heme Fe proteins serves this role in providing the extra electron(s) required for reactivity [1]. We have recently reviewed our studies on mononuclear non-heme Fe enzymes [21]. Here, we focus on the binuclear non-heme Fe enzymes and the new modes of O2 activation associated with utilizing pairs of interacting metal centers in O2 binding and catalysis. The proteins we have studied, and results obtained are summarized in Table 1.
Table 1.
Binuclear non-heme iron enzymes studied using VTVH MCD spectroscopy
Protein | Function | Coord. numbera | Bridges | Ligation | Intermediates |
---|---|---|---|---|---|
Category 1 | |||||
Hemerythrin (Hr) [10, 23, 70, 75] | Reversible O2 binding | 6C/5C | (μ-1,3-COO)2 (μOH) | 5H/1E/1D | FeIIIFeIII–OOH |
Category 2 | |||||
Class Ia ribonucleotide reductase (RNR) [11, 55, 56, 79, 81, 82, 96–103] | Biferrous + Y → biferric + Y· | 4C/5C | (μ-1,3-COO)2 | 2H/3E/1D | P (wt mouse and D84E), P′(W48A), X |
Stearoyl Δ9-desaturase (Δ9D) [12, 35, 54, 63, 77, 85, 104] | Stearoyl-acyl carrier → oleoyl-acyl carrier | 5C/5C (without Stearoyl-ACP); 4C/5C (with Stearoyl-ACP) | (μ-1,3-COO)2 | 2H/4E | P |
Soluble methane monooxygenase (sMMO) [13, 61, 105–115] | CH4 + O2 → CH3OH + H2O | 5C/5C | (μ-1,1-COO) (μ-1,3-COO) | 2H/4E | Hperoxo, Q |
Toluene 4-monooxygenase (T4MO) [17, 51, 116, 117] | Toluene → p-cresol | 5C/5C | (μ-1,1-COO) (μ-1,3-COO) | 2H/4E | [P′(wt), P(T201S)]b [78, 80] |
Ferritin [118–120] | Fe2+ → Fe3+ and O2 → H2O2 (Fe storage) | 5C/5C | μ-1,3-COO | 1H/3E/1D/1Q | P |
Bacterioferritin [58, 121–123] | Fe2+ → Fe3+ and O2 → H2O2 (Fe Storage) | 5C/5C | (μ-1,3-COO)2 | 2H/4E | |
DNA protection during starvation (DPS) proteinsc [124, 125] | Fe2+ → Fe3+ and O2/H2O2 → H2O | 6C | 1H/1D/1E | ||
p-aminobenzoate N-oxygenase (AurF) [24, 25, 59, 126] | p-Amino → p-Nitrobenzoate | 5C/5C | (μ-1,3-COO)2 | 3H/4E | P′ |
Category 3 | |||||
Myo-inositol oxygenase (MIOX) [26, 27, 30–32, 127, 128] | Myo-inositol → d-glucuronate | 6C Fe(II)/6C Fe(II) (without MI); 6C Fe(II)/6C Fe(III) (without MI); 5C Fe(II)/6C Fe(III) (with MI) | (μ-1,3-COO) | 4H/2D | G, H |
Flavin diiron proteins (FDP) [18, 28, 29, 68, 69] | O2/2NO → H2O/N2O | 4C/6C | (μ-1,3-COO) | 4H/2D/1E | {FeNO}7FeII |
Unless otherwise indicated, ligation is for the biferrous state
In related enzyme, toluene/o-xylene monooxygenase ToMO
Only 1 Fe(II) binds with high affinity
The binuclear non-heme Fe proteins mostly have a 2His/4carboxylate ligand set with bridging carboxylates and in some cases an oxygen (OH− or oxo) bridge [22]; however, the reversible O2 binding site in hemerythrin (Hr) has five His ligands [23]. It had been thought that this additional His ligation stabilizes the active site from O2 activation; however, p-aminobenzoate N-oxygenase (AurF), myo-inositol oxygenase (MIOX), and the Flavin diiron proteins (FDPs) have more than two His ligands [24–29]. Most of the binuclear non-heme Fe proteins react from their 2Fe(II) state. However, MIOX is more reactive with O2 from its mixed-valent Fe(II)Fe(III) state [30–32], and the 2Fe(II) state of the FDPs is the only binuclear non-heme Fe enzyme site capable of the reduction of 2NO to N2O [33, 34]. In addition, in some enzymes, the resting 2Fe(II) state is relatively stable in O2 and must be activated by substrate (stearoyl Δ9−-desaturase (Δ9D) [35], arylamine oxygenase of the chloramphenicol biosynthetic pathway (CmlI) [36]) or coupling protein (component B in soluble methane monooxygenase (sMMO) [37, 38] and component D in toluene 4-monooxygenase (T4MO) [16]) binding for reaction with O2. It should also be noted that in the class Ia ribonucleotide reductases (RNRs), the biferrous site reacts with O2 (plus an electron) to generate a Tyr radical that transfers to an adjacent subunit for catalysis in DNA synthesis [11].
Over the years, we have developed variable-temperature, variable-field magnetic circular dichroism (VTVH MCD) spectroscopy in the near-IR (NIR) region as a probe of the geometric and electronic structures of ferrous active sites [1, 39]. In “VTVH MCD of biferrous active sites” of this Perspective, we focus on the new features associated with a binuclear Fe(II) center and in “Structure/function correlations over 2Fe(II) active sites”, we use these to define structure/ function correlations over the three structural types of 2Fe(II) active sites in Table 1. The rest of the Perspective focuses on peroxide-level intermediates in reversible O2 binding and activation for H-atom abstraction and EAS. Spectroscopy coupled to electronic structure calculations provides insight into binding and activating peroxo intermediates by binuclear non-heme Fe enzymes for function.
VTVH MCD of biferrous active sites
As shown in Fig. 1a, a mononuclear high-spin Fe(II) site has two dπ→dσ ligand field (LF) transitions at 10 Dq which for the N/O ligation of non-heme, Fe(II) is ~10,000 cm−1 [1, 39–41]. In a low symmetry protein site, these transitions split in energy dependent on the coordination geometry of the Fe(II). For a six coordinate (6C) LF, there are two transitions at ~10,000 cm−1 split by ~2000 cm−1; for a 5C square pyramidal LF, there are LF transitions at >10,000 and ~5000 cm−1. In a 5C trigonal bipyramidal LF, these transitions shift to below 10,000 cm−1 and below 5000 cm−1 (the latter is often not observed due to instrumental cutoff); for 4C distorted tetrahedral centers, there are only LF transitions in the 5000–7000 cm−1 region. This LF transition splitting is thus a direct probe of Fe(II) coordination. However, these transitions are parity forbidden and in the near-IR spectral region, and are generally too weak relative to the background to observe in absorption spectroscopy for metalloproteins. However, high-spin Fe(II) sites have S = 2 ground states, and, thus, are paramagnetic resulting in these LF transitions being intense in the MCD spectrum at low temperature, due to the C-term selection rule associated with degenerate ground states (see [42–44]). As shown in Fig. 1b, the intensity of these LF transitions in MCD at low temperature increases with increasing magnetic field eventually leveling off (i.e., a saturation effect due to the population of only the lowest magnetic-field-split sublevel of the ground state). A series of isotherms with increasing magnetic field taken over a range of temperatures gives the nested set of saturation magnetization curves for a non-heme Fe(II) site presented in Fig. 1c. The S = 2 ground state is five-fold degenerate in MS which undergo a zero field splitting (ZFS) in a low symmetry protein site as given by the first two terms in the spin Hamiltonian in Eq. (1) [45, 46]. These are further split by an applied magnetic field, as shown in Fig. 1d. The saturation magnetization curves in Fig. 1c can be fit to the ZFS plus Zeeman splitting in Fig. 1d to obtain the spin Hamiltonian parameters (and δ and g|| of the lowest doublet) which in turn (i.e., through in-state spin–orbit coupling) reflect the low symmetry splitting of the 5T2 dπ orbitals in Fig. 1a [47, 48].
Fig. 1.
a LF splitting of a high-spin Fe(II) d6 system with variations in coordination number and geometry. Adapted with permission from [41]. Copyright 2005 Royal Society of Chemistry. b Variable field at low temperature, below 5 K, and c variable-temperature, variable-field MCD spectra of Fe(HB(3,5-iPr2pz)3)(S-C6H4-4-tBu). Reprinted with permission from [40]. Copyright 1998 American Chemical Society. d Ground-state energy-level diagram of a high-spin Fe(II) d6 system with the effects of axial and rhombic zero field, and Zeeman splittings. Left positive ZFS with H perpendicular to z; right negative ZFS with H parallel to z. Adapted with permission from [129]. Copyright 1992 American Chemical Society
Spin Hamiltonian for a non-heme Fe(II) center. D and E are the axial and rhombic ZFS parameters, respectively. β, g, and H are the Bohr magneton, g-factor, and the magnetic-field strength, respectively [45, 46]:
(1) |
Extension of the above to a binuclear non-heme Fe(II) enzyme active site leads to two dπ→dσ LF transitions for each Fe(II) center (4 total), a different ZFS on each Fe(II), and the possibility of exchange coupling between the two Fe(II)s due to the presence of bridging ligation. The magnetic interaction between the two Fe(II)s due to their covalent interactions with the bridging ligands (called superexchange pathways) gives the −2JSA·SB term in Eq. (2) which in the absence of ZFS couples the two S = 2 monomer ground states to form total spin S = 4, 3, 2, 1, 0 states for the binuclear site [S = 0 is the antiferromagnetic (AF) ground state and S = 4 is the ferromagnetic (F) ground state reflecting orthogonal superexchange pathways through the bridge] [49]. For biferrous systems, the exchange and ZFS can be comparable leading to a series of J/D diagrams to describe the energies and wavefunctions of the spin sublevels of the coupled binuclear Fe(II) ground state depending on whether the coupling is AF [J negative in Eq. (2)] or F (J positive) and the sign of the ZFS on each Fe(II) and the relative magnitudes of J and D’s [1].
Spin Hamiltonian for an exchange-coupled (J) binuclear non-heme FeA(II)FeB(II) site [49]:
(2) |
MCD data for two very different sets of bridging ligands found in binuclear non-heme Fe(II) active sites are shown in Fig. 2 top with associated J/D diagrams given at the bottom. For deoxyHr in contrast to the behavior of a mononuclear Fe(II) site where the C-term MCD intensity associated with the paramagnetism increases as 1/T (Fig. 2a top, dashed line) until saturation, deoxyHr has no MCD intensity at low temperature and high field [50]. The absence of MCD intensity at low temperature and high field requires that deoxyHr has a diamagnetic MS = 0 ground state. Increasing the temperature leads to the appearance of and increase in MCD intensity which peaks at ~70 K and then decreases in going to higher temperatures (data points and solid curve fit in Fig. 2a top). This temperature-dependent MCD intensity variation requires the presence of a paramagnetic doublet at ~55 cm−1 above the ground state (insert in Fig. 2a top).
Fig. 2.
a Temperature-dependent MCD intensity of deoxyHr (top) and its associated J/D diagram (bottom). Adapted with permission from [50]. Copyright 1987 American Chemical Society. b VTVH MCD of T4MOH (top) [51] and its associated J/D diagram (bottom) [50]. Adapted with permission from [50] and [51]. Copyright 2008 and 1987 American Chemical Society
The right-hand side of the J/D diagram in Fig. 2a bottom gives the limit of no exchange coupling with positive ZFS on both Fe(II)s. Fitting the MCD data in Fig. 2a using only ZFS would require that |D| > 35 cm−1 in Eq. (2) (with J = 0), which is not physically reasonable. The left-hand side of Fig. 2a bottom gives the AF coupling of the 2Fe(II) with D = 0. Fitting the data in Fig. 2a to this limit would give a −J ~ 25 cm−1, which is comparable to D’s of high-spin Fe(II)s. The middle of the J/D diagram gives the combination of AF J and positive D’s and fits the deoxyHr data with −J = 12 ± 2 cm−1. This antiferromagnetic coupling constant is large for a biferrous site and required that deoxyHr have an OH− bridge capable of significant AF coupling of the two Fe(II) centers.
For the biferrous active site of T4MO, a very different ground-state behavior was observed (Fig. 2b) [51]. As the temperature is decreased, the MCD signal intensity increases consistent with T4MO having the C-term behavior of a paramagnetic ground state. The VTVH MCD data in Fig. 2b, however, show a very rapid saturation behavior for the low-temperature isotherms indicating a very high-effective g value for the ground doublet. For mononuclear Fe(II) sites, g||,effective is close to 8 (dashed curve in Fig. 2b top), while for T4MO (data points and solid curves in Fig. 2b top), the g|| is close to 16, reflecting a ferromagnetically exchange-coupled biferrous ground state. The appropriate J/D diagram is given in Fig. 2b bottom. The center gives the energy-level splitting for a ferromagnetically coupled ground state with a positive D to the right and a negative D to the left. The arrow describes the region that fits the VTVH MCD data and indicates that the biferrous active site of T4MO is weakly ferromagnetically coupled with J ~ 1 cm−1. This ferromagnetic coupling is associated with a μ-1,1 carboxylate bridge at the biferrous active site. Other biferrous active sites with only carboxylate bridges, but bridging in a μ-1,3 mode, show VTVH MCD data indicating weak antiferromagnetic coupling (ground states with MS = 0, 1, or 2 depending on the signs of D and the magnitude of D/J) on the order of −J ~ 1 cm−1 [1, 52].
It should finally be noted that in general, each Fe(II) ligand field is different with different ZFSs. Therefore, each contributes differently to the exchange-coupled sublevels of a 2Fe(II) active site (i.e., to the dimer spin states in the J/D diagrams in Fig. 2 bottom). VTVH MCD data taken at each LF transition energy of a biferrous active site (up to 4 in total) will then, in general, show different behaviors enabling the LF transitions to be correlated to each Fe(II) center, thus defining the coordination environment of each Fe(II) in the coupled binuclear Fe(II) active site [53, 54]. VTVH MCD studies have been performed on the biferrous active sites of all the proteins and enzymes included in Table 1 to define the coordination environment of each Fe(II) and the bridging ligation and their changes with perturbation (substrate binding, etc.). These results are summarized in Table 1 and generalized below to develop structure/function correlations.
Structure/function correlations over 2Fe(II) active sites
Based on VTVH MCD data and crystallography, we divide the proteins and enzymes in Table 1 into three categories based on geometric and electronic structures. All the enzyme active sites in category 2 have similar structural features: open coordination positions on both Fe(II) centers and weak exchange coupling (|J| ≤ 1 cm−1) between the irons. RNR [55, 56], ferritin [57], bacterioferritin [58], and AurF [59] all rapidly react with dioxygen to generate peroxy intermediates. Alternatively, substrate binding in Δ9D and component B(D) binding in sMMO (T4MO) are required for rapid reaction with O2 to generate peroxy intermediates [16, 35, 37, 38]. For the latter enzymes, the VTVH MCD data show that a significant biferrous active-site structural change is associated with the increased O2 reactivity [51, 60–62]. This structure/function correlation is illustrated with the VTVH MCD data for Δ9D in Fig. 3 [60]. The resting 2Fe(II) Δ9D site (Fig. 3a) shows only two LF transitions with temperature and field dependence indicating two Fe(II)s with equivalent negative ZFS and weak AF exchange coupling. This MCD analysis is consistent with the X-ray crystal structure showing two equivalent 5C Fe(II)s with μ-1,3-carboxylate bridges [63]. Figure 3b shows the large change in MCD spectral features associated with substrate [stearoyl bound acyl carrier protein (ACP)] binding to the biferrous enzyme. There are now three LF transitions and the two Fe(II)s which are weakly exchange coupled now have different ZFSs. These data show that an active-site structural change is required for O2 activation to the peroxide level.
Fig. 3.
CD spectra at 5 °C and MCD spectra at 7 T, 5 K (left) and VTVH MCD with fit energy-level diagram in insert (right) of Δ9D in the absence (a) and the presence (b) of stearoyl-ACP substrate Adapted with permission from [60]. Copyright 1999 American Chemical Society
The above-mentioned analysis reflects the idea that, for the active sites of the enzymes in category 2, O2 must bridge the 2Fe(II)s for 2e− reduction to the peroxide level. As shown in Fig. 4 [64], if O2 binds to a single Fe(II) in a biferrous site with only carboxylate bridges, it is reduced by 1e− to superoxide that is AF coupled to the resultant high-spin Fe(III). Importantly, there is no significant e− transfer from the second Fe(II) [labeled Fe(1) in Fig. 4]. The exchange coupling J relates to the superexchange pathways between the Fe centers which give the electronic coupling for electron transfer. The low J values associated with the carboxylate bridges reflect poor superexchange pathways for electron transfer (ET) from Fe(1) to the Fe(2)–O2 complex. As shown in Fig. 4, O2 end-on binding to a single Fe is uphill relative to the biferrous site and O2. Alternatively, if O2 is allowed to bridge between the two Fe centers, it is now reduced by 2e− to a peroxide bridging two Fe(III)s in an exothermic process. The significant energy difference reflects the energetically favored two 2e− reduction of O2 to peroxide (360 vs. −160 meV for 1e− reduction of O2) [65, 66], the ability of the peroxide to form strong donor bridges to each of the two Fe(III)s in a μ-1,2-bridged state (vide infra) and, as developed in [67], and the weak binding of superoxide to a high-spin Fe(III). Thus, there is the geometric requirement that the open coordination position on each Fe(II) is oriented, such that O2 can bridge for its 2e− reduction. There is also an electronic requirement. O2 is a triplet with two half-occupied perpendicular π* orbitals. Each Fe(II) is high-spin d6 with an extra e− in a dπ* orbital, as shown in Fig. 5 for Δ9D, where there is spectral/ structural information available for the reactive biferrous state from Fig. 3b [60]. These β HOMOs on the two Fe(II) s should be oriented, such that one is in the Fe–O2 plane and the other is perpendicular to this plane of a bridging O2 such that there is efficient ET into its two perpendicular π* orbitals.
Fig. 4.
Energy diagram for the binding of O2 to a biferrous site (a) terminal to Fe(2) as superoxide and (b) cis-μ-1,2-bridged to the two Fe centers as peroxide Reprinted with permission from [64]. Copyright 2005 American Chemical Society
Fig. 5.
Orbital interactions for the O2 reaction of Δ9D in the absence (left) and the presence (right) of stearoyl-ACP substrate. The β π* orbitals of O2 are unoccupied, while the dπ orbitals shown contain the high-spin d6 β e−. Adapted with permission from [60]. Copyright 1999 American Chemical Society
MIOX and FDP are placed in a separate category 3, where there is again weak exchange coupling between the two Fe(II)s indicating only μ-1,3-carboxylate bridges with poor superexchange pathways for ET between the two Fe centers. However, now, one or both Fe(II)s is coordinatively saturated (i.e., 6C) [32]. For the FDPs, a 4C Fe(II) rapidly reacts with NO in a 1e− process to form high-spin Fe(III)(S = 5/2)–NO−(S = 1) AF coupled, which is an exothermic reaction [68, 69]. Alternatively, MIOX must be oxidized by 1e− to a mixed-valent Fe(II)Fe(III) state that upon substrate binding to the Fe(III) has an open coordination position on its Fe(II) that does react with O2 to form a superoxide intermediate (G) [32]. The geometric and electronic structural changes that occur in going from the substrate-bound Fe(III)Fe(II) to the substrate-bound Fe(III) Fe(III)O2− state in MIOX that enable its 1e− reduction of O2 to form an Fe(III)-bound superoxide are currently unclear.
Finally, we include Hr by itself in category 1. It has an open coordination position on only 1 Fe(II), yet reversibly reacts with dioxygen to reduce it by 2e− to a hydroperoxide [10]. However, as developed in “VTVH MCD of biferrous active sites”, deoxyHr is different from the other binuclear non-heme Fe enzymes in having strong AF coupling requiring an OH− bridge. As shown in Fig. 6, O2 binding at the open coordination position on one Fe induces a proton-coupled 2e− transfer process, where the μ-OH bridge donates its proton to form the oxo bridge of oxyHr, thereby creating a very efficient superexchange pathway for ET [70]. The electronic structure enabling the reversible O2 binding in Fig. 6 is summarized in the next section.
Fig. 6.
Reversible O2 binding to deoxyHr Reprinted with permission from [70]. Copyright 1999 American Chemical Society
Reversible O2 binding: Hr
As shown in Fig. 6, O2 binding to deoxyHr to form oxyHr involves proton-coupled 2e− transfer to O2 binding at a single Fe center [10]. The geometric and electronic structures of deoxyHr are considered above. The electronic structure of oxyHr is defined by its charge transfer (CT) spectrum which reflects the covalent HOO–Fe(III) and Fe(III)–μO–Fe(III) bonds. These CT transitions overlap each other in energy, but as shown in Fig. 7, could be resolved by polarized single crystal electronic absorption (Abs) spectroscopy on oxyHr [70, 71]. The HOO-Fe(III) CT transitions are polarized perpendicular to the Fe–Fe vector (dashed line in Fig. 7b). Peroxide to Fe(III) CT generally involves intense peroxide to dπ and peroxide to dσ CT (see the next section). However, in oxyHr, there are two sets of two peroxo-to-Fe(III) CT transitions that are all comparably weak in intensity (Fig. 7b, dotted Gaussians). This difference in CT intensity reflects the effect of protonation of the peroxide, which mixes the peroxo π* orbitals and reduces the donor interaction of the peroxide with the Fe(III) which decreases the CT intensity. Thus, electron transfer to Fe should be coupled to proton transfer from the hydroperoxide to the bridge. The μ-oxo-to-Fe(III) CT transitions are dominantly polarized along the Fe–Fe vector (solid line in Fig. 7b). We have shown that the CT transitions of bridging ligands reflect the superexchange pathways responsible for ground-state AF coupling [72, 73]. As observed by comparing Fig. 7b, c (solid curves), the CT transitions of the bent oxo bridge in Hr (125° due to the presence of two additional μ-1,3 carboxylate bridges) are quite different from those of linear oxo bridged biferric dimers. In the latter case, π to π superexchange dominates, while in the bent case, the π to π exchange pathway is limited and instead compensated by π to σ superexchange pathways. For this reason, the J values of oxo bridged biferric complexes are not very dependent on Fe–O–Fe angle [74]. As indicated above, superexchange pathways for AF coupling can be related to electronic coupling, and thus, the orbital interactions involved in ET. This point is developed below.
Fig. 7.
a Electronic absorption spectrum of oxyHr in solution. b Single crystal absorption spectra of oxyHr with parallel (solid) and perpendicular (dashed) polarization. Gaussian deconvolution of the perpendicular absorption (corrected for an oxo CT contribution to the perpendicular polarization due to the Fe–O–Fe angle of 125°) is given as dotted curves. c Single crystal absorption spectra of enH2[(FeHEDTA)2]·6H2O, which has a linear Fe–O–Fe angle, with parallel (solid) and perpendicular (dashed) polarization Adapted with permission from [71, 130] and [70]. Copyright 1978, 1989, and 1999 American Chemical Society
We can interpolate between the electronic structures of oxy and deoxy Hr for insight into this 2e− proton-coupled electron transfer (PCET) process [75]. As shown in Fig. 8a, the elongation of the Fe(1)–(OOH) bond of oxyHr leads to the donation of a ↓ electron from the hydroperoxide π* donor orbital to the dπ orbital on Fe(1) (on left). Associated with this bond elongation, the total energy increases, and the pKa of the hydroperoxide decreases while that of the oxo bridge increases. At an Fe-(OOH) distance of 2.9 Å, the complex has a ferrous(hydroxo-superoxo) electronic structure, and thus, it is isoenergetic to have the proton either on the superoxide or on the oxo bridge (red and blue curves in Fig. 8b). These energetics reflect a potential energy surface with a double minimum (Fig. 8c) that leads to rapid proton tunneling between these free bases and thus a low kinetic isotope effect (KIE) for O2 binding [76]. Associated with the proton transfer from the now hydro-superoxide, as shown in Fig. 8a bottom, an ↑ electron is transferred adiabatically (i.e. continuously) from the HOO–Fe(1) orbital to the dπ orbital on the remote Fe(2). Importantly, while this transfer is a PCET process, it is not an H-atom transfer, as the e− is transferred through the π/σ pathway of the bent oxo bridge. This description of the PCET process then correlates the AF coupling associated with the bent oxo bridge with its effectiveness as a superexchange pathway for ET between the Fe centers.
Fig. 8.
a oxyHr FMO variations involved in the PCET process induced by the elongation of Fe(1)–(OOH) bond. b Potential energy surface (PES) for the elongation of the Fe–(O2) bond with the proton on the O2 (red circles) and with the proton on the oxo bridge (blue squares). c Proton PES as a function of the peroxo-proton distance to the oxo bridge Adapted with permission from [75]. Copyright 1999 American Chemical Society
Peroxy intermediates
Enzymes in category 2 react with O2 to form bridged peroxo-Fe(III)2 intermediates, where the high-spin iron centers are antiferromagnetically coupled. Some of these intermediates (P) display a characteristic peroxo-to-Fe(III) charge transfer band at ~700 nm (ε ~ 1500 M−1cm−1) and Mössbauer parameters of δ ~ 0.66 and ΔEQ ~ 1.5 mm/s [35, 55, 56, 77, 78], while others (P′) lack the absorption feature at ~700 nm and display a reduced Mössbauer isomer shift of δ ~ 0.52 mm/s (Table 1) [59, 79, 80]. The P′ intermediates exhibit electrophilic reactivity in triggering the 1e− oxidation of a nearby tryptophan (W48) to convert to a high-valent Fe(IV)Fe(III) intermediate X in RNR [79] and in performing EAS in ToMO [80]. In contrast, the P intermediates are less active; a chemical rescue experiment with the W48A variants of RNR and an indole electron source showed that P converts to P′ for its reduction [79, 81], and P in Δ9D does not initiate substrate dehydrogenation [35, 77]. Based on the resonance Raman excitation profiles of the P intermediate in W48F/D84E RNR (Fig. 9), P was determined to have a cis-μ-1,2-peroxo bridge as in an equivalent structurally-defined model complex [82, 83]. DFT computational studies to evaluate the spectral and reactivity differences between P and P′ suggested a hydroperoxo (i.e., protonated) structure for P′ (Fig. 10; computational details in supporting information). Protonation of the peroxide in P would result in an increase in the peroxo-to-Fe CT band energy and a decrease in intensity and thus no prominent absorption at ~700 nm, and a diminished electron donation to Fe(III) and thus a decreased Mössbauer isomer shift [84–86].
Fig. 9.
rR excitation profiles of the P intermediate in W48F/D84E RNR (top) and the [Fe(III)2(μ-1,2-O2)(OBz)2{HB(pz′)3}2] complex, where OBz is benzoate and HB(pz′)3 is hydrotris(3,5-diisopropyl-1-pyrazolyl)borate Adapted with permission from [82] and [83]. Copyright 1998 and 2004 American Chemical Society
Fig. 10.
μ-1,2-peroxo and μ-1,2-hydroperoxo structures for P (left) and P′ (right) and their peroxo-σ-antibonding LUMOs (bottom with relative energies)
Moreover, the presence of the protonated peroxide in P′ relative to P can explain P′s enhanced electrophilicity compared to P [84]. This protonation results in a stabilization of the peroxo-σ*-based LUMO that accepts electrons for reductive O–O bond cleavage and EAS (Fig. 10, bottom). Accordingly, P′ in the wt RNR active site is calculated to be capable of W48 oxidation with an exergonic ΔG0 of −4.4 kcal/mol, which results in the formation of an (Fe1(III)-hydroxo)(Fe2(IV) = oxo) initial species that would go on to form intermediate X (Fig. 11a, bottom coordinate). In contrast, the reduction of P by W48 and its subsequent O–O bond cleavage are calculated to have a large barrier and be endergonic with ΔG0 of +5.0 kcal/ mol, leading to an Fe1(III)–μO–(Fe2(IV) = oxo) initial structure.
Fig. 11.
Calculated peroxy reaction coordinates for RNR. a 1-electron reduction of P′ (green) and P (red), b conversion of a μ-1,2-to a μ-1,1-hydroperoxo structure, and c F208 hydroxylation by peroxo (red) and hydroperoxo (green) intermediates
The P′ reactivity in ToMO can also be tested in the RNR active site; the O2 reaction of the W48F/D84E variant of RNR leads to an increase in absorption first at ~700 nm, next at ~500 nm, and finally at ~550 nm; these appear to correlate with the formation of P, then P′, and finally, the Fe(III)2 resting site bound by a nearby hydroxylated F208 residue [87]. In evaluating this reaction, the μ-1,2-hydroperoxo biferric structure in the D84E RNR active site is calculated to be almost isoenergetic to a μ-1,1-hydroperoxo biferric structure with a reasonable energy barrier for interconversion, ΔG‡ of 7.6 kcal/mol (Fig. 11b). This μ-1,1-hydroperoxo structure can oxygenate F208 with a calculated energy barrier of ΔG‡ ~ 17.5 kcal/ mol, consistent with the observed kinetics (2.75×10−5/s; ΔG‡ ~ 22 kcal/mol) for the 500 nm-band decay to form the 550 nm bound hydroxylated product (Fig. 11c bottom reaction coordinate). Alternatively, the corresponding reaction with P is calculated to be highly unlikely with ΔG‡ ~ 36.9 kcal/mol (Fig. 11c top reaction coordinate). This significant reactivity difference with protonation again reflects its effect on the peroxo-σ* orbital which is the frontier molecular orbital (FMO) in these reactions. The cis-μ-1,2 to μ-1,1 hydroperoxo conversion in Fig. 11b is required for this reactivity, as it orients the peroxo-σ* orbital for effective overlap with the aromatic substrate.
Concluding comments
The studies summarized above are now being extended in a number of directions. Nuclear resonance vibrational spectroscopy (NRVS) is being used for the experimental structural determination of the peroxo-level intermediates and the high-valent intermediates X in RNR and Q in sMMO to define geometric and electronic structure contributions to their reactivity [88, 89]. Spectroscopic studies on de novo binuclear Fe(II) proteins with systematic variation in His/ CO2− ligation are being pursued to understand how this ligand variation affects structure and reactivity [90]. In addition, MnFe analogues of 2Fe sites are present in class Ic RNR [91] and the R2lox [92] enzymes. The Mn(III)/ Mn(IV) center can be probed by MCD and the Fe(III) center by NRVS to correlate with the binuclear non-heme Fe enzymes [93]. Finally, a number of new biferrous enzyme classes are emerging (deoxyhypusine hydroxylase (DOHH) [94, 131], urease [95], CmlI [132], etc.) that are accessible to the VTVH MCD/DFT approach presented above to extend Table 1 and develop additional structure/function correlations.
Supplementary Material
Acknowledgments
EIS thanks Prof. J. Martin Bollinger and Prof. Carsten D. Krebs (Penn State U), Prof. Brian G. Fox (U of Wisconsin-Madison), Prof. Donald M. Kurtz Jr. (U of Texas at San Antonio), Prof. John D. Lipscomb (U of Minnesota), Prof. Elizabeth C. Theil (Children’s Hospital Oakland Research Institute and North Carolina State U), Prof. Lawrence Que, Jr. (U of Minnesota), and past graduate students and postdocs for their major contributions to these studies. This work was supported by the National Science Foundation (MCB1404866 to EIS), National Institutes of Health (GM040392 to EIS), C1 Gas Refinery Program through the National Research Foundation of Korea (2015M3D3A1A01064889 to KP), and Korea Institute of Science and Technology Information (KSC-2015-C1-030 to KP).
Footnotes
Electronic supplementary material The online version of this article (doi: 10.1007/s00775-016-1372-9) contains supplementary material, which is available to authorized users.
References
- 1.Solomon EI, Brunold TC, Davis MI, Kemsley JN, Lee SK, Lehnert N, Neese F, Skulan AJ, Yang YS, Zhou J. Chem Rev. 2000;100:235–350. doi: 10.1021/cr9900275. [DOI] [PubMed] [Google Scholar]
- 2.Krebs C, Bollinger JM, Jr, Booker SJ. Curr Opin Chem Biol. 2011;15:291–303. doi: 10.1016/j.cbpa.2011.02.019. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 3.Pootoolal J, Thomas MG, Marshall CG, Neu JM, Hubbard BK, Walsh CT, Wright GD. Proc Natl Acad Sci USA. 2002;99:8962–8967. doi: 10.1073/pnas.102285099. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 4.Ziehl M, He J, Dahse H-M, Hertweck C. Angew Chem Int Ed Engl. 2005;44:1202–1205. doi: 10.1002/anie.200461990. [DOI] [PubMed] [Google Scholar]
- 5.Yang B, Hodgkinson A, Millward BA, Demaine AG. Int J Diabetes Mellit. 2010;2:169–174. [Google Scholar]
- 6.Shao J, Zhou B, Chu B, Yen Y. Curr Cancer Drug Targets. 2006;6:409–431. doi: 10.2174/156800906777723949. [DOI] [PubMed] [Google Scholar]
- 7.Fishman A, Tao Y, Bentley WE, Wood TK. Biotechnol Bioeng. 2004;87:779–790. doi: 10.1002/bit.20185. [DOI] [PubMed] [Google Scholar]
- 8.Schirmer A, Rude MA, Li X, Popova E, del Cardayre SB. Science. 2010;329:559–562. doi: 10.1126/science.1187936. [DOI] [PubMed] [Google Scholar]
- 9.Lee S-W, Keeney DR, Lim D-H, Dispirito AA, Semrau JD. Appl Environ Microbiol. 2006;72:7503–7509. doi: 10.1128/AEM.01604-06. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 10.Stenkamp RE. Chem Rev. 1994;94:715–726. [Google Scholar]
- 11.Stubbe J. Curr Opin Chem Biol. 2003;7:183–188. doi: 10.1016/s1367-5931(03)00025-5. [DOI] [PubMed] [Google Scholar]
- 12.Fox BG, Shanklin J, Somerville C, Munck E. Proc Natl Acad Sci USA. 1993;90:2486–2490. doi: 10.1073/pnas.90.6.2486. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13.Tinberg CE, Lippard SJ. Acc Chem Res. 2011;44:280–288. doi: 10.1021/ar1001473. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 14.Makris TM, Chakrabarti M, Münck E, Lipscomb JD. Proc Natl Acad Sci USA. 2010;107:15391–15396. doi: 10.1073/pnas.1007953107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 15.Austin RN, Chang H-K, Zylstra GJ, Groves JT. J Am Chem Soc. 2000;122:11747–11748. [Google Scholar]
- 16.Lountos GT, Mitchell KH, Studts JM, Fox BG, Orville AM. Biochemistry. 2005;44:7131–7142. doi: 10.1021/bi047459g. [DOI] [PubMed] [Google Scholar]
- 17.Sazinsky MH, Bard J, Di Donato A, Lippard SJ. J Biol Chem. 2004;279:30600–30610. doi: 10.1074/jbc.M400710200. [DOI] [PubMed] [Google Scholar]
- 18.Kurtz DM. Dalton Trans. 2007;37:4115–4121. [Google Scholar]
- 19.Solomon EI, Decker A, Lehnert N. Proc Natl Acad Sci USA. 2003;100:3589–3594. doi: 10.1073/pnas.0336792100. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20.Krebs C, Galonic Fujimori D, Walsh CT, Bollinger JM., Jr Acc Chem Res. 2007;40:484–492. doi: 10.1021/ar700066p. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 21.Solomon EI, Light KM, Liu LV, Srnec M, Wong SD. Acc Chem Res. 2013;46:2725–2739. doi: 10.1021/ar400149m. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 22.Nordlund P, Eklund H. Curr Opin Struct Biol. 1995;5:758–766. doi: 10.1016/0959-440x(95)80008-5. [DOI] [PubMed] [Google Scholar]
- 23.Holmes MA, Le Trong I, Turley S, Sieker LC, Stenkamp RE. J Mol Biol. 1991;218:583–593. doi: 10.1016/0022-2836(91)90703-9. [DOI] [PubMed] [Google Scholar]
- 24.Zocher G, Winkler R, Hertweck C, Schulz GE. J Mol Biol. 2007;373:65–74. doi: 10.1016/j.jmb.2007.06.014. [DOI] [PubMed] [Google Scholar]
- 25.Choi YS, Zhang H, Brunzelle JS, Nair SK, Zhao H. Proc Natl Acad Sci USA. 2008;105:6858–6863. doi: 10.1073/pnas.0712073105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 26.Brown PM, Caradoc-Davies TT, Dickson JM, Cooper GJ, Loomes KM, Baker EN. Proc Natl Acad Sci USA. 2006;103:15032–15037. doi: 10.1073/pnas.0605143103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 27.Brown PM, Caradoc-Davies TT, Dickson JM, Cooper GJ, Loomes KM, Baker EN. Acta Crystallograph Sect F Struct Biol Cryst Commun. 2006;62:811–813. doi: 10.1107/S1744309106028144. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 28.Seedorf H, Hagemeier CH, Shima S, Thauer RK, Warkentin E, Ermler U. FEBS J. 2007;274:1588–1599. doi: 10.1111/j.1742-4658.2007.05706.x. [DOI] [PubMed] [Google Scholar]
- 29.Di Matteo A, Scandurra FM, Testa F, Forte E, Sarti P, Brunori M, Giuffre A. J Biol Chem. 2008;283:4061–4068. doi: 10.1074/jbc.M705605200. Epub 2007 Dec 4012. [DOI] [PubMed] [Google Scholar]
- 30.Bollinger JM, Jr, Diao Y, Matthews ML, Xing G, Krebs C. Dalton Trans. 2009 doi: 10.1039/b811885j. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 31.Xing G, Barr EW, Diao Y, Hoffart LM, Prabhu KS, Arner RJ, Reddy CC, Krebs C, Bollinger JM., Jr Biochemistry. 2006;45:5402–5412. doi: 10.1021/bi0526276. [DOI] [PubMed] [Google Scholar]
- 32.Snyder RA, Bell CB, Diao Y, Krebs C, Bollinger JM, Solomon EI. J Am Chem Soc. 2013;135:15851–15863. doi: 10.1021/ja406635k. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 33.Silaghi-Dumitrescu R, Coulter ED, Das A, Ljungdahl LG, Jameson GNL, Huynh BH, Kurtz DM. Biochem. 2003;42:2806–2815. doi: 10.1021/bi027253k. [DOI] [PubMed] [Google Scholar]
- 34.Fang H, Caranto JD, Mendoza R, Taylor AB, Hart PJ, Kurtz DM. J Biol Inorg Chem. 2012;17:1231–1239. doi: 10.1007/s00775-012-0938-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 35.Broadwater JA, Achim C, Münck E, Fox BG. Biochemistry. 1999;38:12197–12204. doi: 10.1021/bi9914199. [DOI] [PubMed] [Google Scholar]
- 36.Makris TM, Vu VV, Meier KK, Komor AJ, Rivard BS, Münck E, Que L, Lipscomb JD. J Am Chem Soc. 2015;137:1608–1617. doi: 10.1021/ja511649n. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 37.Fox BG, Liu Y, Dege JE, Lipscomb JD. J Biol Chem. 1991;266:540–550. [PubMed] [Google Scholar]
- 38.Liu KE, Valentine AM, Wang D, Huynh BH, Edmondson DE, Salifoglou A, Lippard SJ. J Am Chem Soc. 1995;117:10174–10185. [Google Scholar]
- 39.Solomon EI, Pavel EG, Loeb KE, Campochiaro C. Coord Chem Rev. 1995;144:369–460. [Google Scholar]
- 40.Pavel EG, Kitajima N, Solomon EI. J Am Chem Soc. 1998;120:3949–3962. [Google Scholar]
- 41.Neidig ML, Solomon EI. Chem Commun (Camb) 2005;(47):5843–5863. doi: 10.1039/b510233m. [DOI] [PubMed] [Google Scholar]
- 42.Stephens PJ. J Chem Phys. 1970;52:3489–3516. [Google Scholar]
- 43.Stephens PJ. Annu Rev Phys Chem. 1974;25:201–232. [Google Scholar]
- 44.Stephens PJ. Advances in chemical physics. John Wiley & Sons Inc; New York: 2007. pp. 197–264. [Google Scholar]
- 45.Abragam A, Bleaney B. Electron paramagnetic resonance of transition ions. Clarendon Press; Oxford: 1970. [Google Scholar]
- 46.Bleaney B, Stevens KWH. Rep Prog Phys. 1953;16:108. [Google Scholar]
- 47.Zhang Y, Gebhard MS, Solomon EI. J Am Chem Soc. 1991;113:5162–5175. [Google Scholar]
- 48.Solomon EI, Zhou J, Neese F, Pavel EG. Chem Biol. 1997;4:795–808. doi: 10.1016/s1074-5521(97)90113-7. [DOI] [PubMed] [Google Scholar]
- 49.Moriya T. Phys Rev. 1960;120:91–98. [Google Scholar]
- 50.Reem RC, Solomon EI. J Am Chem Soc. 1987;109:1216–1226. [Google Scholar]
- 51.Schwartz JK, Wei P-P, Mitchell KH, Fox BG, Solomon EI. J Am Chem Soc. 2008;130:7098–7109. doi: 10.1021/ja800654d. [DOI] [PubMed] [Google Scholar]
- 52.Pulver SC, Tong WH, Bollinger JM, Stubbe J, Solomon EI. J Am Chem Soc. 1995;117:12664–12678. [Google Scholar]
- 53.Neese F, Solomon EI. Inorg Chem. 1999;38:1847–1865. doi: 10.1021/ic981264d. [DOI] [PubMed] [Google Scholar]
- 54.Yang YS, Baldwin J, Ley BA, Bollinger JM, Jr, Solomon EI. J Am Chem Soc. 2000;122:8495–8510. [Google Scholar]
- 55.Bollinger JM, Jr, Krebs C, Vicol A, Chen S, Ley BA, Edmondson DE, Huynh BH. J Am Chem Soc. 1998;120:1094–1095. [Google Scholar]
- 56.Yun D, Garcia-Serres R, Chicalese BM, An YH, Huynh BH, Bollinger JM., Jr Biochemistry. 2007;46:1925–1932. doi: 10.1021/bi061717n. [DOI] [PubMed] [Google Scholar]
- 57.Tosha T, Hasan MR, Theil EC. Proc Natl Acad Sci USA. 2008;105:18182–18187. doi: 10.1073/pnas.0805083105. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 58.Macedo S, Romao CV, Mitchell E, Matias PM, Liu MY, Xavier AV, LeGall J, Teixeira M, Lindley P, Carrondo MA. Nat Struct Biol. 2003;10:285–290. doi: 10.1038/nsb909. [DOI] [PubMed] [Google Scholar]
- 59.Korboukh VK, Li N, Barr EW, Bollinger JM, Jr, Krebs C. J Am Chem Soc. 2009;131:13608–13609. doi: 10.1021/ja9064969. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 60.Yang Y-S, Broadwater JA, Pulver SC, Fox BG, Solomon EI. J Am Chem Soc. 1999;121:2770–2783. [Google Scholar]
- 61.Mitić N, Schwartz JK, Brazeau BJ, Lipscomb JD, Solomon EI. Biochemistry. 2008;47:8386–8397. doi: 10.1021/bi800818w. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 62.Pulver SC, Froland WA, Lipscomb JD, Solomon EI. J Am Chem Soc. 1997;119:387–395. [Google Scholar]
- 63.Lindqvist Y, Huang W, Schneider G, Shanklin J. EMBO J. 1996;15:4081–4092. [PMC free article] [PubMed] [Google Scholar]
- 64.Wei P-P, Skulan AJ, Wade H, DeGrado WF, Solomon EI. J Am Chem Soc. 2005;127:16098–16106. doi: 10.1021/ja053661a. [DOI] [PubMed] [Google Scholar]
- 65.Sawyer DT. Oxygen chemistry. Oxford University Press; New York: 1991. [Google Scholar]
- 66.Holm RH, Kennepohl P, Solomon EI. Chem Rev. 1996;96:2239–2314. doi: 10.1021/cr9500390. [DOI] [PubMed] [Google Scholar]
- 67.Schenk G, Pau MY, Solomon EI. J Am Chem Soc. 2004;126:505–515. doi: 10.1021/ja036715u. [DOI] [PubMed] [Google Scholar]
- 68.Hayashi T, Caranto JD, Wampler DA, Kurtz DM, Moënne-Loccoz P. Biochemistry. 2010;49:7040–7049. doi: 10.1021/bi100788y. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 69.Snyder RA, Kurtz DM, Jr, Solomon EI. to be published. [Google Scholar]
- 70.Brunold TC, Solomon EI. J Am Chem Soc. 1999;121:8277–8287. [Google Scholar]
- 71.Reem RC, McCormick JM, Richardson DE, Devlin FJ, Stephens PJ, Musselman RL, Solomon EI. J Am Chem Soc. 1989;111:4688–4704. [Google Scholar]
- 72.Tuczek F, Solomon EI. J Am Chem Soc. 1994;116:6916–6924. [Google Scholar]
- 73.Solomon EI, Lever ABP, editors. Inorganic electronic structure and spectroscopy. John Wiley & Sons Inc; New York: 1999. [Google Scholar]
- 74.Gorun SM, Lippard SJ. Inorg Chem. 1991;30:1625–1630. [Google Scholar]
- 75.Brunold TC, Solomon EI. J Am Chem Soc. 1999;121:8288–8295. [Google Scholar]
- 76.Armstrong GD, Sykes AG. Inorg Chem. 1986;25:3135–3139. [Google Scholar]
- 77.Broadwater JA, Ai J, Loehr TM, Sanders-Loehr J, Fox BG. Biochemistry. 1998;37:14664–14671. doi: 10.1021/bi981839i. [DOI] [PubMed] [Google Scholar]
- 78.Song WJ, Behan RK, Naik SG, Huynh BH, Lippard SJ. J Am Chem Soc. 2009;131:6074–6075. doi: 10.1021/ja9011782. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 79.Saleh L, Krebs C, Ley BA, Naik S, Huynh BH, Bollinger JM., Jr Biochemistry. 2004;43:5953–5964. doi: 10.1021/bi036099e. [DOI] [PubMed] [Google Scholar]
- 80.Murray LJ, Naik SG, Ortillo DO, Garcia-Serres R, Lee JK, Huynh BH, Lippard SJ. J Am Chem Soc. 2007;129:14500–14510. doi: 10.1021/ja076121h. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 81.Saleh L, Kelch BA, Pathickal BA, Baldwin J, Ley BA, Bollinger JM., Jr Biochemistry. 2004;43:5943–5952. doi: 10.1021/bi036098m. [DOI] [PubMed] [Google Scholar]
- 82.Skulan AJ, Brunold TC, Baldwin J, Saleh L, Bollinger JM, Jr, Solomon EI. J Am Chem Soc. 2004;126:8842–8855. doi: 10.1021/ja049106a. [DOI] [PubMed] [Google Scholar]
- 83.Brunold TC, Tamura N, Kitajima N, Moro-oka Y, Solomon EI. J Am Chem Soc. 1998;120:5674–5690. [Google Scholar]
- 84.Jensen KP, Bell CB, 3rd, Clay MD, Solomon EI. J Am Chem Soc. 2009;131:12155–12171. doi: 10.1021/ja809983g. [DOI] [PubMed] [Google Scholar]
- 85.Srnec M, Rokob TA, Schwartz JK, Kwak Y, Rulíšek L, Solomon EI. Inorg Chem. 2012;51:2806–2820. doi: 10.1021/ic2018067. [DOI] [PubMed] [Google Scholar]
- 86.Chalupský J, Rokob TA, Kurashige Y, Yanai T, Solomon EI, Rulíšek L, Srnec M. J Am Chem Soc. 2014;136:15977–15991. doi: 10.1021/ja506934k. [DOI] [PubMed] [Google Scholar]
- 87.Baldwin J, Voegtli WC, Khidekel N, Moënne-Loccoz P, Krebs C, Pereira AS, Ley BA, Huynh BH, Loehr TM, Riggs-Gelasco PJ, Rosenzweig AC, Bollinger JM., Jr J Am Chem Soc. 2001;123:7017–7030. doi: 10.1021/ja002114g. [DOI] [PubMed] [Google Scholar]
- 88.Park K, Tsugawa T, Furutachi H, Kwak Y, Liu LV, Wong SD, Yoda Y, Kobayashi Y, Saito M, Kurokuzu M, Seto M, Suzuki M, Solomon EI. Angew Chem Int Ed Engl. 2013;52:1294–1298. doi: 10.1002/anie.201208240. [DOI] [PubMed] [Google Scholar]
- 89.Park K, Bell CB, Liu LV, Wang D, Xue G, Kwak Y, Wong SD, Light KM, Zhao J, Alp EE, Yoda Y, Saito M, Kobayashi Y, Ohta T, Seto M, Que L, Solomon EI. Proc Natl Acad Sci USA. 2013;110:6275–6280. doi: 10.1073/pnas.1304238110. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 90.Reig AJ, Pires MM, Snyder RA, Wu Y, Jo H, Kulp DW, Butch SE, Calhoun JR, Szyperski TG, Solomon EI, DeGrado WF. Nat Chem. 2012;4:900–906. doi: 10.1038/nchem.1454. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 91.Jiang W, Yun D, Saleh L, Barr EW, Xing G, Hoffart LM, Maslak M-A, Krebs C, Bollinger JM. Science. 2007;316:1188–1191. doi: 10.1126/science.1141179. [DOI] [PubMed] [Google Scholar]
- 92.Andersson CS, Högbom M. Proc Natl Acad Sci USA. 2009;106:5633–5638. doi: 10.1073/pnas.0812971106. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 93.Kwak Y, Jiang W, Dassama LMK, Park K, Bell CB, Liu LV, Wong SD, Saito M, Kobayashi Y, Kitao S, Seto M, Yoda Y, Alp EE, Zhao J, Bollinger JM, Krebs C, Solomon EI. J Am Chem Soc. 2013;135:17573–17584. doi: 10.1021/ja409510d. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 94.Vu VV, Emerson JP, Martinho M, Kim YS, Munck E, Park MH, Que L., Jr Proc Natl Acad Sci USA. 2009;106:14814–14819. doi: 10.1073/pnas.0904553106. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 95.Carter EL, Tronrud DE, Taber SR, Karplus PA, Hausinger RP. Proc Natl Acad Sci USA. 2011;108:13095–13099. doi: 10.1073/pnas.1106915108. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 96.Wei P-P, Skulan AJ, Mitić N, Yang YS, Saleh L, Bollinger JM, Jr, Solomon EI. J Am Chem Soc. 2004;126:3777–3788. doi: 10.1021/ja0374731. [DOI] [PubMed] [Google Scholar]
- 97.Sturgeon BE, Burdi D, Chen S, Huynh BH, Edmondson DE, Stubbe J, Hoffman BM. J Am Chem Soc. 1996;118:7551–7557. [Google Scholar]
- 98.Shanmugam M, Doan PE, Lees NS, Stubbe J, Hoffman BM. J Am Chem Soc. 2009;131:3370–3376. doi: 10.1021/ja809223s. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 99.Mitic N, Saleh L, Schenk G, Bollinger JM, Jr, Solomon EI. J Am Chem Soc. 2003;125:11200–11201. doi: 10.1021/ja036556e. [DOI] [PubMed] [Google Scholar]
- 100.Mitić N, Clay MD, Saleh L, Bollinger JM, Jr, Solomon EI. J Am Chem Soc. 2007;129:9049–9065. doi: 10.1021/ja070909i. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 101.Riggs-Gelasco P, Shu L, Chen S, Burdi D, Huynh BH, Que L, Jr, Stubbe J. J Am Chem Soc. 1998;120:849–860. [Google Scholar]
- 102.Dassama LMK, Silakov A, Krest CM, Calixto JC, Krebs C, Bollinger JM, Green MT. J Am Chem Soc. 2013;135:16758–16761. doi: 10.1021/ja407438p. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 103.Doan PE, Shanmugam M, Stubbe J, Hoffman BM. J Am Chem Soc. 2015;137:15558–15566. doi: 10.1021/jacs.5b10763. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 104.Moënne-Loccoz P, Baldwin J, Ley BA, Loehr TM, Bollinger JM., Jr Biochemistry. 1998;37:14659–14663. doi: 10.1021/bi981838q. [DOI] [PubMed] [Google Scholar]
- 105.Whittington DA, Lippard SJ. J Am Chem Soc. 2001;123:827–838. doi: 10.1021/ja003240n. [DOI] [PubMed] [Google Scholar]
- 106.DeWitt JG, Bentsen JG, Rosenzweig AC, Hedman B, Green J, Pilkington S, Papaefthymiou GC, Dalton H, Hodgson KO, Lippard SJ. J Am Chem Soc. 1991;113:9219–9235. [Google Scholar]
- 107.Elango N, Radhakrishnan R, Froland WA, Wallar BJ, Earhart CA, Lipscomb JD, Ohlendorf DH. Protein Sci. 1997;6:556–568. doi: 10.1002/pro.5560060305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 108.Lee SJ, McCormick MS, Lippard SJ, Cho U-S. Nature. 2013;494:380–384. doi: 10.1038/nature11880. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 109.Brazeau BJ, Lipscomb JD. Biochemistry. 2000;39:13503–13515. doi: 10.1021/bi001473l. [DOI] [PubMed] [Google Scholar]
- 110.Shu L, Nesheim JC, Kauffmann K, Münck E, Lipscomb JD, Que L., Jr Science. 1997;275:515–518. doi: 10.1126/science.275.5299.515. [DOI] [PubMed] [Google Scholar]
- 111.Banerjee R, Proshlyakov Y, Lipscomb JD, Proshlyakov DA. Nature. 2015;518:431–434. doi: 10.1038/nature14160. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 112.Valentine AM, Stahl SS, Lippard SJ. J Am Chem Soc. 1999;121:3876–3887. [Google Scholar]
- 113.Tinberg CE, Lippard SJ. Biochemistry. 2010;49:7902–7912. doi: 10.1021/bi1009375. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 114.Lee SK, Lipscomb JD. Biochemistry. 1999;38:4423–4432. doi: 10.1021/bi982712w. [DOI] [PubMed] [Google Scholar]
- 115.Tinberg CE, Lippard SJ. Biochemistry. 2009;48:12145–12158. doi: 10.1021/bi901672n. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 116.McCormick MS, Sazinsky MH, Condon KL, Lippard SJ. J Am Chem Soc. 2006;128:15108–15110. doi: 10.1021/ja064837r. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 117.Skjeldal L, Peterson FC, Doreleijers JF, Moe LA, Pikus JD, Westler WM, Markley JL, Volkman BF, Fox BG. J Biol Inorg Chem. 2004;9:945–953. doi: 10.1007/s00775-004-0594-4. [DOI] [PubMed] [Google Scholar]
- 118.Ha Y, Shi D, Small GW, Theil EC, Allewell NM. J Biol Inorg Chem. 1999;4:243–256. doi: 10.1007/s007750050310. [DOI] [PubMed] [Google Scholar]
- 119.Schwartz JK, Liu XS, Tosha T, Theil EC, Solomon EI. J Am Chem Soc. 2008;130:9441–9450. doi: 10.1021/ja801251q. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 120.Kwak Y, Schwartz JK, Haldar S, Behera RK, Tosha T, Theil EC, Solomon EI. Biochemistry. 2014;53:473–482. doi: 10.1021/bi4013726. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 121.Crow A, Lawson TL, Lewin A, Moore GR, Brun NEL. J Am Chem Soc. 2009;131:6808–6813. doi: 10.1021/ja8093444. [DOI] [PubMed] [Google Scholar]
- 122.van Eerde A, Wolterink-van Loo S, van der Oost J, Dijkstra BW. Protein Struct Commun. 2006;F62:1061–1066. doi: 10.1107/S1744309106039583. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 123.Kwak Y, Schwartz JK, Huang VW, Boice E, Kurtz DM, Solomon EI. Biochemistry. 2015;54:7010–7018. doi: 10.1021/acs.biochem.5b01033. [DOI] [PubMed] [Google Scholar]
- 124.Schwartz JK, Liu XS, Tosha T, Diebold A, Theil EC, Solomon EI. Biochemistry. 2010;49:10516–10525. doi: 10.1021/bi101346c. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 125.Pesek J, Büchler R, Albrecht R, Boland W, Zeth K. J Biol Chem. 2011;286:34872–34882. doi: 10.1074/jbc.M111.246108. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 126.Li N, Korboukh VK, Krebs C, Bollinger JM. Proc Natl Acad Sci USA. 2010;107:15722–15727. doi: 10.1073/pnas.1002785107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 127.Xing G, Hoffart LM, Diao Y, Prabhu KS, Arner RJ, Reddy CC, Krebs C, Bollinger JM., Jr Biochemistry. 2006;45:5393–5401. doi: 10.1021/bi0519607. [DOI] [PubMed] [Google Scholar]
- 128.Xing G, Diao Y, Hoffart LM, Barr EW, Prabhu KS, Arner RJ, Reddy CC, Krebs C, Bollinger JM., Jr Proc Natl Acad Sci USA. 2006;103:6130–6135. doi: 10.1073/pnas.0508473103. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 129.Solomon EI, Zhang Y. Acc Chem Res. 1992;25:343–352. [Google Scholar]
- 130.Gay RR, Solomon EI. J Am Chem Soc. 1978;100:1972–1973. [Google Scholar]
- 131.Jasniewski AJ, Engstrom LM, Vu VV, Park MH, Que L., Jr J Biol Inorg Chem. 2016 doi: 10.1007/s00775-016-1373-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 132.Knoot CJ, Kovaleva EG, Lipscomb JD. J Biol Inorg Chem. 2016 doi: 10.1007/s00775-016-1363-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
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