Jeffrey W. Pavlik

Hydrosulfide (HS-) coordination in iron porphyrinates.

Recent reports of potential physiological roles of hydrogen sulfide have prompted interest in heme-sulfide interactions. Heme-H(2)S and/or heme-HS(-) interactions could potentially occur during endogenous production, transport, signaling events, and catabolism of H(2)S. We have investigated the interaction of the hydrosulfide ion (HS(-)) with iron porphyrinates. UV-vis spectral studies show the formation of [Fe(Por)(SH)](-), [Fe(Por)(SH)(2)](2-), and the mixed-ligand species [Fe(Por)(Im)(SH)](-). UV-vis binding studies of [Fe(OEP)] and [Fe(T-p-OMePP)] (OEP = octaethylporphyrinate; T-p-OMePP = tetra-p-methoxyphenylporphyrinate) with HS(-) allowed for calculation of the formation constants and extinction coefficients of mono- and bis-HS(-) complexes. We report the synthesis of the first HS(-)-bound iron(II) porphyrin compounds, [Na(222)][Fe(OEP)(SH)].0.5C(6)H(6) and [Na(222)][Fe(T-p-OMePP)(SH)].C(6)H(5)Cl (222 = Kryptofix-222). Characterization by single-crystal X-ray analysis, mass spectrometry, and Mossbauer and IR spectroscopy is all consistent with that of known sulfur-bound high-spin iron(II) compounds. The Fe-S distances of 2.3929(5) and 2.3887(13) A are longer than all reported values of [Fe(II)(Por)(SR)](-) species. An analysis of the porphyrin nonplanarity for these derivatives and for all five-coordinate high-spin iron(II) porphyrinate derivatives with an axial anion ligand is presented. In our hands, attempts to synthesize iron(III) HS(-) derivatives led to iron(II) species.

Probing Vibrational Anisotropy with Nuclear Resonance Vibrational Spectroscopy

One of the important issues of nitrosyl (nitric oxide, NO) iron porphyrinate derivatives (hemes) is to develop a detailed understanding of the molecular basis for selectivity (recognition) between the diatomic ligands NO, CO and O2. The sensing of these gaseous molecules is predominantly by heme-based proteins,[1] and heme- and heme protein-diatomic interactions continues to be an active area of research.[2] In Nature, NO is discriminated from O2 quite efficiently by a number of systems;[3]–[5]most prominent are those of soluble guanalyl cyclase and the NO sensing protein of Clostridium botulinum. Conformational changes in the protein imparted upon ligand binding is a plausible explanation for such differentiation.[6, 7] Very low frequency doming modes, sometimes referred to as reactive modes,[8] may facilitate binding and release of diatomic molecules.[9],[10] Infrared and resonance Raman spectroscopy have provided insight into the interplay of structure and function of heme active sites[11] However, these techniques have some inherent limitations, especially in the low frequency regime where mode assignment is hampered by weak signal, spectral congestion and low sensitivity to isotopic substitution.[8] Also, the directionality of iron within particular vibrations, a potential mechanistic indicator, cannot easily be determined. Our recent studies of the nitrosyl (NO) derivatives of iron porphyrinates have emphasized interesting aspects of nitrosyl dynamics of both the five- and six-coordinate NO species,[8],[12]–[16] including Nuclear Resonance Vibrational Spectroscopy (NRVS, sometimes termed Nuclear Inelastic Scattering, NIS) studies.[8],[15]–[19]

New Perspectives on Iron–Ligand Vibrations of Oxyheme Complexes

We report our studies of the vibrational dynamics of iron for three imidazole-ligated oxyheme derivatives that mimic the active sites of histidine-ligated heme proteins complexed with dioxygen. The experimental vibrational data are obtained from nuclear resonance vibrational spectroscopy (NRVS) measurements conducted on both powder samples and oriented single crystals, and which includes several in-plane (ip) and out-of-plane (oop) measurements. Vibrational spectral assignments have been made through a combination of the oriented sample spectra and predictions based on density functional theory (DFT) calculations. The two Fe-O(2) modes that have been previously observed by resonance Raman spectroscopy in heme proteins are clearly shown to be very strongly mixed and are not simply either a bending or stretching mode. In addition, a third Fe-O(2) mode, not previously reported, has been identified. The long-sought Fe-Im stretch, not observed in resonance Raman spectra, has been identified and compared with the frequencies observed for the analogous CO and NO species. The studies also suggest that the in-plane iron motion is anisotropic and is controlled by the orientation of the Fe-O(2) group and not sensitive to the in-plane Fe-N(p) bonds and/or imidazole orientations.

Vibrational Probes and Determinants of the S = 0 ⇌ S = 2 Spin Crossover in Five-Coordinate [Fe(TPP)(CN)]−

The low-frequency vibrational characterization of the spin-crossover complex, five-coordinate cyano(tetraphenylporphyrinato)iron(II), [Fe(TPP)(CN)](-), is reported. Nuclear resonance vibrational spectroscopy has been used to measure all low-frequency vibrations involving iron at several temperatures; this yields vibrational spectra of both the low- (S = 0) and high-spin (S = 2) states. Multitemperature oriented single-crystal measurements facilitate assignments of the vibrational character of all modes and are consistent with the DFT-predicted spectra. The availability of the entire iron vibrational spectrum allows for the complete correlation of the modes between the two spin states. These data demonstrate that not only do the frequencies of the vibrations shift to lower values for the high-spin species as would be expected owing to the weaker bonds in the high-spin state, but also the mixing of iron modes with ligand modes changes substantially. Diagrams illustrating the changing character of the modes and their correlation are given. The reduced iron-ligand frequencies are the primary factor in the entropic stabilization of the high-spin state responsible for the spin crossover.

Anisotropic Iron Motion in Nitrosyl Iron Porphyrinates:Natural and Synthetic Hemes

The vibrational spectra of two five-coordinate nitrosyl iron porphyrinates, [Fe(OEP)(NO)] (OEP = dianion of 2,3,7,8,12,13,17,18-octaethylporphyrin) and [Fe(DPIX)(NO)] (DPIX = deuteroporphyrin IX), have been studied by oriented single-crystal nuclear resonance vibrational spectroscopy. Single crystals (both are in the triclinic crystal system) were oriented to give vibrational spectra perpendicular to the porphyrin plane. Additionally, two orthogonal in-plane measurements that were also either perpendicular or parallel to the projection of the FeNO plane onto the porphyrin plane yield the complete set of vibrations with iron motion. In addition to cleanly enabling the assignment of the FeNO bending and stretching modes, the measurements reveal that the two in-plane spectra from the parallel and perpendicular in-plane directions for both compounds have substantial differences. The assignment of these in-plane vibrations were aided by density functional theory predictions. The differences in the two in-plane directions result from the strongly bonded axial NO ligand. The direction of the in-plane iron motion is thus found to be largely parallel and perpendicular to the projection of the FeNO plane on the porphyrin plane. These axial ligand effects on the in-plane iron motion are related to the strength of the axial ligand-to-iron bond.

Probing Heme Vibrational Anisotropy: An Imidazole Orientation Effect?

The complete iron vibrational spectrum of the five-coordinate high-spin complex [Fe(OEP)(2-MeHIm)], where OEP = octaethylporphyrinato and 2-MeHIm = 2-methylimidazole, has been obtained by oriented single-crystal nuclear resonance vibrational spectroscopy (NRVS) data. Measurements have been made in three orthogonal directions, which provides quantitative information for all iron motion. These experimental data, buttressed by density functional theory (DFT) calculations, have been used to define the effects of the axial ligand orientation. Although the axial imidazole removes the degeneracy in the in-plane vibrations, the imidazole orientation does not appear to control the direction of the in-plane iron motion. This is in contrast to the effect of the imidazolate ligand, as defined by DFT calculations, which does have substantial effects on the direction of the in-plane iron motion. The axial NO ligand has been found to have the strongest orientational effect (Angew. Chem., Int. Ed., 2010, 49, 4400). Thus the strength of the directional properties are in the order NO > imidazolate > imidazole, consistent with the varying strength of the Fe-ligand bond.

3D Motions of Iron in Six-Coordinate {FeNO}(7) Hemes by Nuclear Resonance Vibration Spectroscopy.

The vibrational spectrum of a six-coordinate nitrosyl iron porphyrinate, monoclinic [Fe(TpFPP)(1-MeIm)(NO)] (TpFPP=tetra-para-fluorophenylporphyrin; 1-MeIm=1-methylimidazole), has been studied by oriented single-crystal nuclear resonance vibrational spectroscopy (NRVS). The crystal was oriented to give spectra perpendicular to the porphyrin plane and two in-plane spectra perpendicular or parallel to the projection of the FeNO plane. These enable assignment of the FeNO bending and stretching modes. The measurements reveal that the two in-plane spectra have substantial differences that result from the strongly bonded axial NO ligand. The direction of the in-plane iron motion is found to be largely parallel and perpendicular to the projection of the bent FeNO on the porphyrin plane. The out-of-plane Fe-N-O stretching and bending modes are strongly mixed with each other, as well as with porphyrin ligand modes. The stretch is mixed with v50 as was also observed for dioxygen complexes. The frequency of the assigned stretching mode of eight Fe-X-O (X=N, C, and O) complexes is correlated with the Fe-XO bond lengths. The nature of highest frequency band at ≈560 cm(-1) has also been examined in two additional new derivatives. Previously assigned as the Fe-NO stretch (by resonance Raman), it is better described as the bend, as the motion of the central nitrogen atom of the FeNO group is very large. There is significant mixing of this mode. The results emphasize the importance of mode mixing; the extent of mixing must be related to the peripheral phenyl substituents.