An activity-based fluorescent sensor for the detection of the phenol sulfotransferase SULT1A1 in living cells.

Human phenol sulfotransferases mediate the transfer of a sulfuryl moiety from the activated sulfate donor PAPS to hydroxy-containing substrates, altering substrate solubility and charge to affect phase II metabolism and cell signaling. Here, we present the development, computational modeling, in vitro enzymology, and biological application of STS-3, an activity-based fluorescent sensor for the SULT1A1 isoform.

Biomimetic Reactivity of Oxygen-Derived Manganese and Iron Porphyrinoid Complexes.

Heme proteins utilize the heme cofactor, an iron porphyrin, to perform a diverse range of reactions including dioxygen binding and transport, electron transfer, and oxidation/oxygenations. These reactions share several key metalloporphyrin intermediates, typically derived from dioxygen and its congeners such as hydrogen peroxide. These species are composed of metal-dioxygen, metal-superoxo, metal-peroxo, and metal-oxo adducts. A wide variety of synthetic metalloporphyrinoid complexes have been synthesized to generate and stabilize these intermediates. These complexes have been studied to determine the spectroscopic features, structures, and reactivities of such species in controlled and well-defined environments. In this Review, we summarize recent findings on the reactivity of these species with common porphyrinoid scaffolds employed for biomimetic studies. The proposed mechanisms of action are emphasized. This Review is organized by structural type of metal-oxygen intermediate and broken into subsections based on the metal (manganese and iron) and porphyrinoid ligand (porphyrin, corrole, and corrolazine).

High-Valent Manganese-Oxo Valence Tautomers and the Influence of Lewis/Brönsted Acids on C-H Bond Cleavage.

The addition of Lewis or Brönsted acids (LA = Zn(OTf)2, B(C6F5)3, HBArF, TFA) to the high-valent manganese-oxo complex MnV(O)(TBP8Cz) results in the stabilization of a valence tautomer MnIV(O-LA)(TBP8Cz•+). The ZnII and B(C6F5)3 complexes were characterized by manganese K-edge X-ray absorption spectroscopy (XAS). The position of the edge energies and the intensities of the pre-edge (1s to 3d) peaks confirm that the Mn ion is in the +4 oxidation state. Fitting of the extended X-ray absorption fine structure (EXAFS) region reveals 4 N/O ligands at Mn-Nave = 1.89 Å and a fifth N/O ligand at 1.61 Å, corresponding to the terminal oxo ligand. This Mn-O bond length is elongated compared to the MnV(O) starting material (Mn-O = 1.55 Å). The reactivity of MnIV(O-LA)(TBP8Cz•+) toward C-H substrates was examined, and it was found that H• abstraction from C-H bonds occurs in a 1:1 stoichiometry, giving a MnIV complex and the dehydrogenated organic product. The rates of C-H cleavage are accelerated for the MnIV(O-LA)(TBP8Cz•+) valence tautomer as compared to the MnV(O) valence tautomer when LA = ZnII, B(C6F5)3, and HBArF, whereas for LA = TFA, the C-H cleavage rate is slightly slower than when compared to MnV(O). A large, nonclassical kinetic isotope effect of kH/kD = 25-27 was observed for LA = B(C6F5)3 and HBArF, indicating that H-atom transfer (HAT) is the rate-limiting step in the C-H cleavage reaction and implicating a potential tunneling mechanism for HAT. The reactivity of MnIV(O-LA)(TBP8Cz•+) toward C-H bonds depends on the strength of the Lewis acid. The HAT reactivity is compared with the analogous corrole complex MnIV(O-H)(tpfc•+) recently reported (J. Am. Chem. Soc. 2015, 137, 14481-14487).

The Influence of Peripheral Substituent Modification on P(V), Mn(III), and Mn(V)(O) Corrolazines: X-ray Crystallography, Electrochemical and Spectroscopic Properties, and HAT and OAT Reactivities.

The influence of remote peripheral substitution on the physicochemical properties and reactivity of phosphorus and manganese corrolazine (Cz) complexes was examined. The substitution of p-MeO for p-t-Bu groups on the eight phenyl substituents of the ß-carbon atoms of the Cz ring led to changes in UV-vis transitions and redox potentials for each of the complexes. The oxygen atom transfer (OAT) and hydrogen atom transfer (HAT) reactivity of the Mn(V)(O) complexes was also influenced by p-MeO substitution. The OAT reactivity of Mn(V)(O)(MeOP8Cz) (MeOP8Cz = octakis(p-methoxyphenyl)corrolazinato(3-)) with triarylphosphine (PAr3) substrates led to second-order rate constants from 10.2(5) to 3.1(2) × 10(4) M(-1) s(-1). These rates of OAT are slower than those seen for Mn(V)(O)(TBP8Cz) (TBP8Cz = octakis(p-tert-butylphenyl)corrolazinato(3-)). A Hammett study involving para-substituted PAr3 substrates reveals a Hammett ρ-value for Mn(V)(O)(MeOP8Cz) that is more negative than that observed for Mn(V)(O)(TBP8Cz), consistent with a less electrophilic Mn center. The HAT reactivity of Mn(V)(O)(MeOP8Cz) with C-H substrates was examined and revealed second-order rate constants from 6.8(5) × 10(-5) to 1.70(2) × 10(-1) M(-1) s(-1). The rate constants varied with the C-H bond strength of the substrate. Slightly faster HAT rates with C-H substrates were observed with Mn(V)(O)(MeOP8Cz) compared to Mn(V)(O)(TBP8Cz), indicating that the basicity of the putative [Mn(IV)(O)](-) intermediate likely compensates for the more negative redox potential in the driving force for HAT. In addition, the complete, large-scale synthesis of the para-phenyl-substituted porphyrazines RP8PzH2 (R = p-tert-butylphenyl (TB), p-methoxyphenyl (MeO), and p-isopropylphenyl) and corrolazines RP8CzH3 (TBP8CzH3 and MeOP8CzH3) is presented. The crystal structures of the monoprotonated, metal-free corrolazine [(TBP8CzH3)(H)](+)[BArF](-), P(V)(OMe)2(MeOP8Cz), and Mn(III)(MeOP8Cz)(MeOH) are presented. This work provides the first insights into the influence of electronic substituent effects on the corrolazine periphery.

A Balancing Act: Stability versus Reactivity of Mn(O) Complexes.

A large class of heme and non-heme metalloenzymes utilize O2 or its derivatives (e.g., H2O2) to generate high-valent metal-oxo intermediates for performing challenging and selective oxidations. Due to their reactive nature, these intermediates are often short-lived and very difficult to characterize. Synthetic chemists have sought to prepare analogous metal-oxo complexes with ligands that impart enough stability to allow for their characterization and an examination of their inherent reactivity. The challenge in designing these molecules is to achieve a balance between their stability, which should allow for their in situ characterization or isolation, and their reactivity, in which they can still participate in interesting chemical transformations. This Account focuses on our recent efforts to generate and stabilize high-valent manganese-oxo porphyrinoid complexes and tune their reactivity in the oxidation of organic substrates. Dioxygen can be used to generate a high-valent Mn(V)(O) corrolazine (Mn(V)(O)(TBP8Cz)) by irradiation of Mn(III)(TBP8Cz) with visible light in the presence of a C-H substrate. Quantitative formation of the Mn(V)(O) complex occurs with concomitant selective hydroxylation of the benzylic substrate hexamethylbenzene. Addition of a strong H(+) donor converted this light/O2/substrate reaction from a stoichiometric to a catalytic process with modest turnovers. The addition of H(+) likely activates a transient Mn(V)(O) complex to achieve turnover, whereas in the absence of H(+), the Mn(V)(O) complex is an unreactive "dead-end" complex. Addition of anionic donors to the Mn(V)(O) complex also leads to enhanced reactivity, with a large increase in the rate of two-electron oxygen atom transfer (OAT) to thioether substrates. Spectroscopic characterization (Mn K-edge X-ray absorption and resonance Raman spectroscopies) revealed that the anionic donors (X(-)) bind to the Mn(V) ion to form six-coordinate [Mn(V)(O)(X)](-) complexes. An unusual "V-shaped" Hammett plot for the oxidation of para-substituted thioanisole derivatives suggested that six-coordinate [Mn(V)(O)(X)](-) complexes can act as both electrophiles and nucleophiles, depending on the nature of the substrate. Oxidation of the Mn(V)(O) corrolazine resulted in the in situ generation of a Mn(V)(O) π-radical cation complex, [Mn(V)(O)(TBP8Cz(•+))](+), which exhibited more than a 100-fold rate increase in the oxidation of thioethers. The addition of Lewis acids (LA; Zn(II), B(C6F5)3) to the closed-shell, diamagnetic Mn(V)(O)(TBP8Cz) stabilized a paramagnetic valence tautomer Mn(IV)(O)(TBP8Cz(•+))(LA), which was characterized as a second π-radical cation complex by NMR, EPR, UV-vis, and high resolution cold spray ionization MS. The Mn(IV)(O)(TBP8Cz(•+))(LA) complexes are able to abstract H(•) from phenols and exhibit a rate enhancement of up to ∼100-fold over the parent Mn(V)(O) valence tautomer. In contrast, a large decrease in rate is observed for OAT for the Mn(IV)(O)(TBP8Cz(•+))(LA) complexes. The rate enhancement for hydrogen atom transfer (HAT) may derive from the higher redox potential for the π-radical cation complex, while the large rate decrease seen for OAT may come from a decrease in electrophilicity for an Mn(IV)(O) versus Mn(V)(O) complex.

Mn(V)(O) versus Cr(V)(O) Porphyrinoid Complexes: Structural Characterization and Implications for Basicity Controlling H-Atom Abstraction.

Isomorphous crystals of Mn(V)(O) and Cr(V)(O) corrolazines were characterized by single crystal X-ray diffraction. Reactivity studies with H atom donors and separated PCET reagents show a dramatic difference in H atom abstracting abilities for these two complexes. The implied large difference in driving force is opposite the trend in redox potentials, indicating that basicity is a key factor in determining the striking difference in reactivity for two metal-oxo species in identical ligand environments.

Strong Inhibition of O-Atom Transfer Reactivity for Mn(IV)(O)(π-Radical-Cation)(Lewis Acid) versus Mn(V)(O) Porphyrinoid Complexes.

The oxygen atom transfer (OAT) reactivity of two valence tautomers of a Mn(V)(O) porphyrinoid complex was compared. The OAT kinetics of Mn(V)(O)(TBP8Cz) (TBP8Cz = octakis(p-tert-butylphenyl)corrolazinato(3-)) reacting with a series of triarylphosphine (PAr3) substrates were monitored by stopped-flow UV-vis spectroscopy, and revealed second-order rate constants ranging from 16(1) to 1.43(6) × 10(4) M(-1) s(-1). Characterization of the OAT transition state analogues Mn(III)(OPPh3)(TBP8Cz) and Mn(III)(OP(o-tolyl)3)(TBP8Cz) was carried out by single-crystal X-ray diffraction (XRD). A valence tautomer of the closed-shell Mn(V)(O)(TBP8Cz) can be stabilized by the addition of Lewis and Brønsted acids, resulting in the open-shell Mn(IV)(O)(TBP8Cz(•+)):LA (LA = Zn(II), B(C6F5)3, H(+)) complexes. These Mn(IV)(O)(π-radical-cation) derivatives exhibit dramatically inhibited rates of OAT with the PAr3 substrates (k = 8.5(2) × 10(-3) - 8.7 M(-1) s(-1)), contrasting the previously observed rate increase of H-atom transfer (HAT) for Mn(IV)(O)(TBP8Cz(•+)):LA with phenols. A Hammett analysis showed that the OAT reactivity for Mn(IV)(O)(TBP8Cz(•+)):LA is influenced by the Lewis acid strength. Spectral redox titration of Mn(IV)(O)(TBP8Cz(•+)):Zn(II) gives Ered = 0.69 V vs SCE, which is nearly +700 mV above its valence tautomer Mn(V)(O)(TBP8Cz) (Ered = -0.05 V). These data suggest that the two-electron electrophilicity of the Mn(O) valence tautomers dominate OAT reactivity and do not follow the trend in one-electron redox potentials, which appear to dominate HAT reactivity. This study provides new fundamental insights regarding the relative OAT and HAT reactivity of valence tautomers such as M(V)(O)(porph) versus M(IV)(O)(porph(•+)) (M = Mn or Fe) found in heme enzymes.

Light-driven, proton-controlled, catalytic aerobic C-H oxidation mediated by a Mn(III) porphyrinoid complex.

The visible light-driven, catalytic aerobic oxidation of benzylic C-H bonds was mediated by a Mn(III) corrolazine complex. To achieve catalytic turnovers, a strict selective requirement for the addition of protons was established. The resting state of the catalyst was unambiguously characterized by X-ray diffraction as [Mn(III)(H2O)(TBP8Cz(H))](+), in which a single, remote site on the ligand is protonated. If two remote sites are protonated, however, reactivity with O2 is shut down. Spectroscopic methods revealed that the related Mn(V)(O) complex is also protonated at the same remote site at -60 °C, but undergoes valence tautomerization upon warming.

Mechanistic elucidation of the stepwise formation of a tetranuclear manganese pinned butterfly cluster via N-N bond cleavage, hydrogen atom transfer, and cluster rearrangement.

A mechanistic pathway for the formation of the structurally characterized manganese-amide-hydrazide pinned butterfly complex, Mn4(µ3-PhN-NPh-κ(3)N,N')2(µ-PhN-NPh-κ(2)-N,N')(µ-NHPh)2L4 (L = THF, py), is proposed and supported by the use of labeling studies, kinetic measurements, kinetic competition experiments, kinetic isotope effects, and hydrogen atom transfer reagent substitution, and via the isolation and characterization of intermediates using X-ray diffraction and electron paramagnetic resonance spectroscopy. The data support a formation mechanism whereby bis[bis(trimethylsilyl)amido]manganese(II) (Mn(NR2)2, where R = SiMe3) reacts with N,N'-diphenylhydrazine (PhNHNHPh) via initial proton transfer, followed by reductive N-N bond cleavage to form a long-lived Mn(IV) imido multinuclear complex. Coordinating solvents activate this cluster for abstraction of hydrogen atoms from an additional equivalent of PhNHNHPh resulting in a Mn(II)phenylamido dimer, Mn2(µ-NHPh)2(NR2)2L2. This dimeric complex further assembles in fast steps with two additional equivalents of PhNHNHPh replacing the terminal silylamido ligands with η(1)-hydrazine ligands to give a dimeric Mn2(µ-NHPh)2(PhN-NHPh)2L4 intermediate, and finally, the addition of two additional equivalents of Mn(NR2)2 and PhNHNHPh gives the pinned butterfly cluster.

Oxygen-atom transfer reactivity of axially ligated Mn(V)-oxo complexes: evidence for enhanced electrophilic and nucleophilic pathways.

Addition of anionic donors to the manganese(V)-oxo corrolazine complex Mn(V)(O)(TBP8Cz) has a dramatic influence on oxygen-atom transfer (OAT) reactivity with thioether substrates. The six-coordinate anionic [Mn(V)(O)(TBP8Cz)(X)](-) complexes (X = F(-), N3(-), OCN(-)) exhibit a ∼5 cm(-1) downshift of the Mn-O vibrational mode relative to the parent Mn(V)(O)(TBP8Cz) complex as seen by resonance Raman spectroscopy. Product analysis shows that the oxidation of thioether substrates gives sulfoxide product, consistent with single OAT. A wide range of OAT reactivity is seen for the different axial ligands, with the following trend determined from a comparison of their second-order rate constants for sulfoxidation: five-coordinate ≈ thiocyanate ≈ nitrate < cyanate < azide < fluoride ≪ cyanide. This trend correlates with DFT calculations on the binding of the axial donors to the parent Mn(V)(O)(TBP8Cz) complex. A Hammett study was performed with p-X-C6H4SCH3 derivatives and [Mn(V)(O)(TBP8Cz)(X)](-) (X = CN(-) or F(-)) as the oxidant, and unusual "V-shaped" Hammett plots were obtained. These results are rationalized based upon a change in mechanism that hinges on the ability of the [Mn(V)(O)(TBP8Cz)(X)](-) complexes to function as either an electrophilic or weak nucleophilic oxidant depending upon the nature of the para-X substituents. For comparison, the one-electron-oxidized cationic Mn(V)(O)(TBP8Cz(•+)) complex yielded a linear Hammett relationship for all substrates (ρ = -1.40), consistent with a straightforward electrophilic mechanism. This study provides new, fundamental insights regarding the influence of axial donors on high-valent Mn(V)(O) porphyrinoid complexes.

Activation of a high-valent manganese-oxo complex by a nonmetallic Lewis acid.

The reaction of a manganese(V)-oxo porphyrinoid complex with the Lewis acid B(C6F5)3 leads to reversible stabilization of the valence tautomer Mn(IV)(O)(π-radical cation). The latter complex, in combination with B(C6F5)3, reacts with ArO-H substrates via formal hydrogen-atom transfer and exhibits dramatically increased reaction rates over the Mn(V)(O) starting material.

Valence tautomerism in a high-valent manganese-oxo porphyrinoid complex induced by a Lewis acid.

Addition of the Lewis acid Zn(2+) to (TBP(8)Cz)Mn(V)(O) induces valence tautomerization, resulting in the formation of [(TBP(8)Cz(+•))Mn(IV)(O)-Zn(2+)]. This new species was characterized by UV-vis, EPR, the Evans method, and (1)H NMR and supported by DFT calculations. Removal of Zn(2+) quantitatively restores the starting material. Electron-transfer and hydrogen-atom-transfer reactions are strongly influenced by the presence of Zn(2+).

Synthesis of tetranuclear, four-coordinate manganese clusters with "pinned butterfly" geometry formed by metal-mediated N-N bond cleavage in diphenylhydrazine.

The preparation of four-coordinate tetramanganese-amide-hydrazide clusters is described. Reaction of Mn(NR(2))(2) (R = SiMe(3)) with N,N'-diphenylhydrazine resulted in the formation of a black intermediary mixture that converted to a four-coordinate tetranuclear "pinned butterfly" cluster, Mn(4)(µ(3)-N(2)Ph(2))(2)(µ-N(2)Ph(2))(µ-NHPh)(2)(THF)(4). This compound was isolated in ~90% yield and identified by single-crystal X-ray diffraction analysis. In pyridine, the THF ligands were replaced, giving the pyridyl complex Mn(4)(µ(3)-N(2)Ph(2))(2)(µ-N(2)Ph(2))(µ-NHPh)(2)(py)(4). Charge counting considerations indicate that the clusters had gained two protons and two electrons in addition to the formative fragments. Isolation of the black mixture was achieved by extraction techniques from a reaction with a decreased loading of hydrazine run at low temperatures with decreased solvent polarity. The black mixture was characterized by FT-IR, UV-vis, and (1)H NMR spectroscopy. In addition, an isolable, colorless dimer, Mn(2)(µ-NHPh)(2)(NR(2))(2)(THF)(2), was present in the mixture and identified by single-crystal X-ray diffraction. These intermediates are discussed in light of possible mechanisms for formation of the tetranuclear cluster.