Nuclear Quadrupole Interaction Parameters Research Articles

Electronic Structure Study of Certain Rhizoferrin Analogs and Its Ferric-Ion Complexes

We have used first-principles electronic structure methods to examine the ferric-ion binding specificity of rhizoferrin analogs: (a) the homo-rhizoferrin and nor-rhizoferrin that fall in the category of variable length diamine bridge; (b) monodesoxy-rhizoferrin and didesoxy-rhizoferrin having different functional groups in the citric-acid moieties of rhizoferrin, and (c) cyclized dehydration product of rhizoferrin, named imido-rhizoferrin. A particular focus is the effect of deprotonation of hydroxyl and carboxyl groups on binding energy of ferric-ion complexes of rhizoferrin-analogs has been investigated. In addition, we have calculated Nuclear Magnetic Resonance properties, such as chemical shifts for13C and 1H, and indirect spin-spin couplings for methylene protons, and compared our results with available experimental data. The nuclear quadrupole interaction parameters will be discussed within the context of charge distributions and electric field gradients. Our study provides a structural basis for ferric-ion chelation by rhizoferrin analogs for these biomolecules, for which there exist no experimentally determined structures. Argonne National Laboratory is a US DOE Science Laboratory operated under contract no. DE-AC02-06CH11357 by UChicago Argonne, LLC. The research was performed, in part, at Argonne National Laboratory as a research participant in the Visiting Faculty Program. The program is administered by Argonne's Division of Communication, Education, and Public Affairs (CEPA) with funding provided by the U.S. Department of Energy.

First-principles electronic structure study of rhizoferrin and its Fe(III) complexes

We have determined the structure and coordination chemistry of rhizoferrin (Rf), which is a particular type of siderophore, and its Fe(III) complexes using density functional theory calculations. Our results show that the Fe(III) ion binds in an octahedral coordination, with a low-spin (S = 1/2) charge-neutral chiral complex having the largest binding energy of the investigated complexes. We have also calculated nuclear magnetic resonance parameters, such as chemical shifts for (1)H and (13)C, and indirect nuclear spin-spin couplings for (1)H-(1)H and (13)C-(1)H in free Rf and in a low-spin neutral Rf metal complex, as well as nuclear quadrupole interaction parameters, such as asymmetry parameter and nuclear quadrupole coupling constants for (14)N. Our calculated values for the chemical shifts for free Rf are in excellent agreement with experimental data while the calculated NMR parameters for Fe(III) complexes are predictions for future experimental work.

Structure and Conformation of Rhizoferrin and its Fe3+ Complexes

Rhizoferrin is a simple but efficient siderophore and is potentially important for medical applications. The aim of this investigation is to determine the molecular structure of free rhizoferrin as well as its Fe3+ complexes, and to understand the mechanisms of cation (Fe3+)-binding specificity of rhizoferrin, using first-principle methods. The optimized molecular structures, and the calculated Nuclear Magnetic Resonance properties, such as chemical shifts for 13C and 1H and indirect dipole dipolecouplings for 1H, will be presented and compared with the experimental data. The nuclear quadrupole interaction parameters will be discussed as an attempt to understand the charge distribution and electric field gradient. Possible influence of deprotonation of citrate carboxylate groups on Fe3+-rhizoferrin complex formation will be discussed. This is, to the best of our knowledge, the first theoretical study of rhizoferrin and its metal complexes, and it provides a critical insight and guidance to the structural basis of metal chelation by rhizoferrin.

11B and 23Na solid-state NMR and density functional theory studies of electric field gradients at boron sites in ulexite

Nuclear magnetic resonance (NMR) parameters of 11B in borates and borosilicates, unlike those of many other nuclei such as 29Si and 27Al, vary only over limited ranges and have been thought to be insensitive to local structural environments. High-resolution NMR spectroscopy at high (14 T) and ultrahigh (21 T) fields yield precise 11B and 23Na NMR parameters for ulexite, which contains the pentaborate polyanion ([B5O6(OH)6]3−) as the fundamental building block (FBB). These NMR parameters are compared with ab initio theoretical calculations as implemented in WIEN2K, including optimization of the ulexite structure, determination of the electric field gradients (EFG) and consequently the nuclear quadrupole interaction (QI) parameters at the five distinct B sites, and calculations of the density of states (DOS). These calculations show that the magnitudes and signs of the EFG for [3]B and [4]B are determined by multiple factors, including the electron distributions in the B 2pz orbitals and their interactions with Ca-3p/O-2s orbitals. Most importantly, the calculated B 2pz orbitals at all B sites in ulexite are predominantly affected by the atoms within the fundamental building block, resulting in the insensitivity of the 11B QI parameters to the weak interunit interactions among FBB. Calculations with the water molecules removed from the ulexite structure provide further support for the strong intraunit interactions in FBB as a cause for the poor sensitivity of 11B NMR parameters to local structural environments, including hydrogen bonding, in borates.

Hafnium oxide thin films studied by time differential perturbed angular correlations

We report on the study of hafnium oxide thin films grown by pulsed laser deposition at various partial oxygen pressures by Time Differential Perturbed Angular Correlations using the nuclear probe 181Hf(β−)181Ta to determine the nuclear quadrupole interaction (NQI), and by x-ray diffraction. The samples were neutron activated and measured at room temperature as received as well as after annealing in air. All spectra exhibited two to three inequivalent probe sites, even after annealing. At 0.3 mbar oxygen partial pressure and annealing for 5 hs at 1073 K the majority (88%) of the sites exhibited NQI parameters as reported for the bulk monoclinic phase [ωQ = 125.4(2) Mrad/s, η = 0.335(5)]. We can exclude amorphous as well as cubic and tetragonal hafnium oxide phases in the annealed samples. There was no indication of room-temperature ferromagnetism.

Pulsed EPR investigations of the Mo(V) centers of the R55Q and R55M variants of sulfite dehydrogenase from Starkeya novella

Continuous-wave and pulsed electron paramagnetic resonance (EPR) spectroscopy have been used to characterize two variants of bacterial sulfite dehydrogenase (SDH) from Starkeya novella in which the conserved active-site arginine residue (R55) is replaced by a neutral amino acid residue. Substitution by the hydrophobic methionine residue (SDH(R55M)) has essentially no effect on the pH dependence of the EPR properties of the Mo(V) center, even though the X-ray structure of this variant shows that the methionine residue is rotated away from the Mo center and a sulfate anion is present in the active-site pocket (Bailey et al. in J Biol Chem 284:2053-2063, 2009). For SDH(R55M) only the high-pH form is observed, and samples prepared in H(2)(17)O-enriched buffer show essentially the same (17)O hyperfine interaction and nuclear quadrupole interaction parameters as SDH(WT) enzyme. However, the pH dependence of the EPR spectra of SDH(R55Q), in which the positively charged arginine is replaced by the neutral hydrophilic glutamine, differs significantly from that of SDH(WT). For SDH(R55Q) the blocked form with bound sulfate is generated at low pH, as verified by (33)S couplings observed upon reduction with (33)S-labeled sulfite. This observation of bound sulfate for SDH(R55Q) supports our previous hypothesis that sulfite-oxidizing enzymes can exhibit multiple pathways for electron transfer and product release (Emesh et al. in Biochemistry 48:2156-2163, 2009). At pH > or = 8 the high-pH form dominates for SDH(R55Q).

Direct Detection and Characterization of Chloride in the Active Site of the Low-pH Form of Sulfite Oxidase Using Electron Spin Echo Envelope Modulation Spectroscopy, Isotopic Labeling, and Density Functional Theory Calculations

Electron spin echo envelope modulation (ESEEM) investigations were carried out on samples of the low-pH (lpH) form of vertebrate sulfite oxidase (SO) prepared with (35)Cl- and (37)Cl-enriched buffers, as well as with buffer containing the natural abundance of Cl isotopes. The isotope-related changes observed in the ESEEM spectra provide direct and unequivocal evidence that Cl(-) is located in close proximity to the Mo(V) center of lpH SO. The measured isotropic hyperfine interaction constant of about 4 MHz ((35)Cl) suggests that the Cl(-) ion is either weakly coordinated to Mo(V) at its otherwise vacant axial position, trans to the oxo ligand, or is hydrogen-bonded to the equatorial exchangeable OH ligand. Scalar relativistic all-electron density functional theory (DFT) calculations of the hyperfine and nuclear quadrupole interaction parameters, along with steric and energetic arguments, strongly support the possibility that Cl(-) is hydrogen-bonded to the equatorial OH ligand rather than being directly coordinated to the Mo(V).

209Bi NQR studies of low-temperature spin–lattice relaxation in Bi 4Ge 3O 12 single crystal

Unusual influence of weak external magnetic fields on quadrupole spin–spin relaxation was earlier found for Bi 4Ge 3O 12 which exhibited, like some other compounds of composition Ba kBi lA mO n (A=Al, B, Ge, Br, Cl), a dramatic increase in the 209Bi NQR line intensities upon switching the magnetic field. The measuring of the nuclear quadrupole interaction parameters gave an evidence for the presence in the compound of local magnetic fields of the order of 20–30 G. The existence of several low-lying crystal field levels for the p-electrons can be admitted in Bi 4Ge 3O 12 which contributes to the anomalies in the 209Bi NQR spectra of this crystal. The present work was carried out to identify the contribution of such paramagnetic centers to the nuclear spin − lattice relaxation. This contribution resulted in the low-temperature minimum on the curve of the nuclear spin–lattice relaxation rate T 1 − 1 vs. temperature, measured for the transition Δ m = 3 / 2 − 5 / 2 in the 209Bi NQR spectrum of the Bi 4Ge 3O 12 single crystal.

Understanding of nuclear quadrupole interactions of 35 Cl, 79 Br and 129 I and binding energies of solid halogens at first-principles level

This paper deals with the understanding at a first-principles level of the nuclear quadrupole interaction (NQI) parameters of solid chlorine, bromine and iodine as well as the intermolecular binding of these molecules in the solid. The electronic structure investigations that we have carried out to study these properties of the solid halogens are based on the Hartree-Fock Cluster approach using the Roothaan variational procedure with electron correlation effects included using many-body perturbation theory with the empty orbitals used in the perturbation theory investigations for the excited states. The results of our investigations provide good agreement with the measured NQI parameters primarily from the Hartree-Fock one electron wave-functions with many-body effects making minor contributions. The binding (dissociation) energies for the molecules with the solid state environment on the other hand arises from intermolecular many body effects identified as the Van der Waals attraction with one-electron Hartree-Fock contribution being repulsive in nature.

Pulsed EPR Investigations of Systems Modeling Molybdenum Enzymes: Hyperfine and Quadrupole Parameters of Oxo-17O in [Mo17O(SPh)4]-

Ka band ESEEM spectroscopy was used to determine the hyperfine (hfi) and nuclear quadrupole (nqi) interaction parameters for the oxo-17O ligand in [Mo 17O(SPh)4]-, a spectroscopic model of the oxo-Mo(V) centers of enzymes. The isotropic hfi constant of 6.5 MHz found for the oxo-17O is much smaller than the values of approximately 20-40 MHz typical for the 17O nucleus of an equatorial OH(2) ligand in molybdenum enzymes. The 17O nqi parameter (e2qQ/h = 1.45 MHz, eta approximately = 0) is the first to be obtained for an oxo group in a metal complex. The parameters of the oxo-17O ligand, as well as other magnetic resonance parameters of [Mo 17O(SPh)4]- predicted by quasi-relativistic DFT calculations, were in good agreement with those obtained in experiment. From the electronic structure of the complex revealed by DFT, it follows that the SOMO is almost entirely molybdenum d(xy) and sulfur p, while the spin density on the oxo-17O is negative, determined by spin polarization mechanisms. The results of this work will enable direct experimental identification of the oxo ligand in a variety of chemical and biological systems.

Study of muon magnetic hyperfine and nuclear quadrupole interactions and easy axis in the organic ferromagnet p-Cl-Ph-CHN-TEMPO

The first-principles Hartree–Fock cluster procedure has been used for studying the electronic structure of the organic ferromagnetic p-Cl-Ph-CHN-TEMPO (4-(p-chlorobenzylideneamino)-2,2,6,6-tetramethylpiperidin-1-yloxyl) radical system with and without positive muon or muonium. Theory is able to provide quantitative information about the trapping sites for muon and muonium and the associated muon hyperfine interaction parameters. Using this information, two sites, one with muonium trapped near oxygen in the electronic singlet state of the muonium–radical pair system involved and the other with muon trapped near chlorine, are identified as likely candidates for the source of the experimentally observed μSR frequency of 3.2 MHz from zero-field measurement. The easy axis is obtained in both cases as the b-axis in the monoclinic lattice in agreement with the results of our magnetic dipole–dipole interaction studies using the magnetic moment distributions in the ferromagnetic state in the absence of muon and muonium. Nuclear quadrupole interaction parameters for nuclei adjacent to the trapped muon and muonium have been evaluated and their role in distinguishing between alternate trapping sites is discussed.

Analysis of correlation patterns in hyperfine sublevel correlation spectroscopy ofS=1/2,I=3/2 systems

The properties of two-dimensional four-pulse electron spin echo envelope modulation spectra of nuclei with nuclear spinI=3/2 in disordered systems are discussed by means of calculated lithium hyperfine sublevel correlation spectra. Since a case of small coupling is treated, the multifrequency transition components appear only in the first quadrant of the two-dimensional spectra. The influence of the hyperfine and nuclear quadrupole interaction parameters on the cross peak ridges in the two-dimensional powder pattern is analyzed. Furthermore the possibility to determine the magnitude of the dipolar hyperfine and the nuclear quadrupole coupling from the cross peak line shapes is investigated. In this context the potential of those spectral features representing the so-called multiquantum transitions for the interpretation of the spectra is demonstrated.

Theory of 14N and 17O Nuclear Quadrupole Interactions in the Single Amino Acids Occurring in the Protein Chain of Cytochrome c

The understanding of electron transport in proteins based on a novel technique involving muon spin rotation (μSR)measurements is a topic of great current interest. The technique, which involves study of spin relaxation of a positive muon (μ+) trapped in amino acids in protein chains due to the fluctuating magnetic field that the moving electron produces, is based on the premise that the electron is generated by ionization of a muonium (Mu) which was trapped at the same site as the μ+ left behind. In attempting to test this premise from first-principles for the Cytochrome c (Cyt c) system in which recent μSR measurements have been made, we have carried out Hartree- Fock investigations of the electronic structures of the bare amino acids and amino acids with + and Mu trapped at the oxygen of the C=O group common to all amino acids. With the aim that the comparison of theoretically predicted experimental nuclear quadrupole interaction (NQI) parameters will provide a useful test of the electron distribution in the amino acids of Cyt c, we present results for the nuclear quadrupole coupling constants (e2qQ) and asymmetry parameters (η) for the bare amino acids and the amino acids with trapped μ+ and Mu. The trends in 2 and for 14N and 17O between the various amino acids, as well as the changes in these parameters in the presence of μ+ and Mu are being analyzed. It would be helpful to have experimental data for e2qQ and to η compare with our predictions for the amino acids as they occur in vitro in polycrystalline Cyt c in which the SR measurements have been carried out. It is also hoped that the μSR technique will be able to provide experimental data on e2qQ and for the 14N and 17O nuclei to compare with our predictions

Temperature Dependence of 2H Nuclear Quadrupole Interaction in Very Short Hydrogen Bonds in Some Organic Acidic Salt Crystals

The 2H nuclear quadrupole interaction parameters, e2Qq/h and η, are closely related to the shape of the potential energy surface at hydrogen bonds and depend sensitively on their geometry. We measured the temperature dependence of the 2H NMR spectra of the crystalline acidic salts KDCO3, KD cetylenedicarboxylate, RbD acetylenedicarboxylate, and KD maleate, which contain very short O-D···O type hydrogen bonds. The temperature coefficient decreases with increase in the O···O distance in the hydrogen bond. Ab inito molecular orbital calculations of the electric field gradient tensor based on the temperature dependent structure of each crystal indicate that thermal expansion of the hydrogen bond geometry is not responsible for this tendency. Ab initio calculations also predict that a fictitious off center shift of the hydrogen position in a symmetric hydrogen bond causes very high e2 Qq/h values. This suggests that low energy vibrational excitation may be responsible for large positive d (e2 Qq/h)/dT values in symmetric hydrogen bonds.

ESEEM studies of succinate:ubiquinone reductase from Paracoccus denitrificans.

Electron spin-echo envelope modulation (ESEEM) spectroscopy has been performed in order to obtain structural information about the environment of the reduced [2Fe-2S] cluster (S-1 center), the oxidized [3Fe-4S] cluster (S-3 center), and the flavin semiquinone radical in purified succinate:ubiquinone reductase from Paracoccus denitrificans. Spectral simulations of the ESEEM data from the reduced [2Fe-2S] yielded nuclear quadrupole interaction parameters that are indicative of peptide nitrogens. We also observed a weak interaction between the oxidized [3Fe-4S] cluster and a peptide 14N. There was no evidence for coordination of any of the Fe atoms to 14N atoms of imidazole rings. The ESEEM data from the flavin semiquinone radical were more complicated. Here, evidence was obtained for interactions between the unpaired electron and only the two nitrogen atoms in the flavin ring.

Effect of Intermolecular Hydrogen Bonding on the Nuclear Quadrupole Interaction in Imidazole and its Derivatives as Studied by ab initio Molecular Orbital Calculations

Ab initio Hartree-Fock molecular orbital calculations were applied to the crystalline imidazole and its derivatives in order to examine systematically the effect of possible N-H---N type hydrogen bond-ing on the nuclear quadrupole interaction parameters in these materials. The nitrogen quadrupole coupling constant (QCC) and the asymmetry parameter (η) of the electric field gradient (EFG) were found to depend strongly on the size of the molecular clusters, from single molecule, to dimer, trimer and to the infinite molecular chain, i.e., crystalline state, implying that the intermolecular N-H -N hydrogen bond affects significantly the electronic structure of imidazole molecule. A certain correla-tion between the QCC of 14N and the N-H bond distance R was also found and interpreted on the basis of the molecular orbital theory. However, we found that the value of the calculated EFG at the hy-drogen position of the N-H group, or the corresponding QCC value of 2 H, increases drastically as R-3 when R is shorter than about 0.1 nm, due probably to the inapplicability of the Gaussian basis sets to the very short chemical bond as revealed in the actual imidazole derivatives. We suggested that the ob-served N-H distances in imidazole derivatives should be re-examined.

Atomic charges in RBa2Cu4O8 lattices, determined by comparison of experimental and calculated nuclear quadrupole interaction parameters

Nuclear quadrupole interaction parameters have been determined for the rare-earth and copper sites of the RBa2Cu4O8 lattices (R = Sm, Y, Er) by 67Cu(67Zn) and 67Ga(67Zn) emission Mössbauer spectroscopy. The lattice electric field gradient tensors have been calculated for all sites of the above lattices using the point charge approximation. A comparison of the measured and calculated parameters has shown that in the RBa2Cu4O8 lattices holes are predominantly confined to the oxygen chain sublattices.

Crystallographic, Electrochemical, and Pulsed EPR Study of Copper(II) Polyimidazole Complexes Relevant to the Metal Sites of Copper Proteins(,).

Copper(II) complexes of the following polyimidazole ligands have been synthesized: bis(imidazol-2-yl)methane (BIM), bis(imidazol-2-yl) ketone (BIK), 4-(imidazol-4-ylmethyl)-2-(imidazol-2-ylmethyl)imidazole (TRIM), bis[4-(imidazol-4-ylmethyl)imidazol-2-yl]methane (TIM), and bis[4-(imidazol-4-ylmethyl)imidazol-2-yl] ketone (TIK). Their crystal structures have been determined using X-ray diffraction. [Cu(ClO(4))(2)(BIM)(2)], 1, belongs to the triclinic space group P&onemacr; system, a = 7.161(4) Å, b = 7.986(6) Å, c = 9.865(3) Å, alpha = 76.73(5) degrees, beta = 71.18(3) degrees, gamma = 76.44(5) degrees, Z = 1, T = 291 K; R = 0.035, R(w) = 0.036 for 1668 reflections; Cu-N = 1.998(3) and 2.001(2) Å, Cu-O = 2.574(4) Å, in a tetragonal geometry. [Cu(BIK)(2)](ClO(4))(2), 2, belongs to the monoclinic space group C2/c system, a = 9.029(3) Å, b = 12.497(2)Å, c = 19.197(2) Å, beta = 94.59(2) degrees, Z = 4, T = 291 K; R = 0.056, R(w) = 0.061 for 1052 reflections; Cu-N = 1.961(7) and 1.954(7) Å, in a distorted tetrahedral geometry. [CuCl(TRIM)(CH(3)OH)]Cl, 6, belongs to the monoclinic space group P2(1)/n system, a = 14.192(5) Å, b = 13.832(5) Å, c = 7.913(3) Å, beta = 90.55(4) degrees, Z = 4, T = 291 K; R = 0.062, R(w) = 0.057 for 1377 reflections; Cu-N = 1.987(7), 2.007(7) and 2.007(6) Å, Cu-O = 2.521(7) Å, Cu-Cl = 2.298(2) Å, in a square pyramidal geometry. [Cu(ClO(4))(TIM)](ClO(4)), 4, belongs to the triclinic space group P&onemacr; system, a = 9.604(4) Å, b = 11.508(6) Å, c = 12.003(8) Å, alpha = 58.79(4) degrees, beta = 94.59(2) degrees, gamma = 67.43(3) degrees, Z = 2, T = 291 K; R = 0.057, R(w) = 0.062 for 2084 reflections; Cu-N = 1.985(7), 1.964(7), 1.967(7), and 1.966(7) Å, Cu-O = 2.553(8) Å, in a distorted square pyramidal geometry. [CuCl(TIK)](ClO(4)), 7, belongs to the triclinic space group P&onemacr; system, a = 7.432(3) Å, b = 12.573(3) Å, c = 12.945(2) Å, alpha = 114.94(4) degrees, beta = 92.46(2) degrees, gamma = 103.49(3) degrees, Z = 2, T = 291 K; R = 0.043, R(w) = 0.049 for 2305 reflections; Cu-N = 1.984(5), 1.989(5), 2.012(5), and 1.979(5) Å, Cu-Cl = 2.796(2) Å, in a distorted bipyramidal geometry. In methanol solution, the perchlorato complexes 1, 2, Cu(TRIM)(ClO(4))(2) (3), 4, and Cu(TIK)(ClO(4))(2) (5) exhibited redox potentials from -215 to +284 mV vs NHE together with a visible absorption from 604 to 728 nm. Electron spin-echo envelope modulation (ESEEM) spectroscopy data, particularly the nuclear quadrupole interaction (NQI) parameters e(2)qQ and eta of the remote nitrogen (N1H), were analyzed and interpreted according to the model devised by Jiang et al. (J. Am. Chem. Soc. 1990, 112, 9035) with reference to Cu(HIm)(4)(ClO(4))(2). The results are the following: (i) C2 substitution of the imidazole ring, next to the remote nitrogen (1, 2) decreases the asymmetry parameter eta to ca. 0.75 compared to 1.00 for Cu(HIm)(4)(ClO(4))(2); this effect of C2 substitution on the symmetry of the electric field gradient at N1H appears similar for both the electron-donating methylene substituents (1) and the electron-withdrawing carbonyl group (2). (ii) The electron-donating or -withdrawing properties of the substituent are reflected by the variation of the e(2)qQ parameter, increasing from 1.43 to 1.75 MHz (1) or decreasing to 1.38 MHz (2), and by the nu(+) transition shifting toward higher frequencies from 1.49 to 1.65 MHz (1, 3, 4) or to lower frequencies to 1.29 MHz (2, 5). The use of the eta and nu(+) parameters to assign the Ndelta vs Nepsilon coordination of histidine to the metal and to detect modified histidine in copper-binding proteins is discussed.

2H and 195 Pt NMR Studies of Molecular and Electron Spin Dynamics in Paramagnetic [Cu(H2O)6][PtCl 6

Abstract The temperature dependences of 2H and 195Pt NMR spectra and the spin-lattice relaxation time T1 were measured for [Cu(H2O)6][PtCl6]. From the simulation of 2H NMR spectra, the jump rate of 180° flips of the water molecules (k), the nuclear quadrupole interaction parameters (e2Qq/h, ƞ) and the electron-nucleon dipolar interaction parameter (vD) were obtained. By measuring 2H T1, k was estimated in the temperature range where the spectrum is insensitive to the motion of the water molecules. Above the phase transition temperature, the pre-exponential factor k0 = 8 x1011 s-1 and the activation energy Ea = 15 kJmol-1 for 180° flips of the water molecules were obtained from the spectral simulation and T1. 195Pt NMR spectra showed an axially symmetric and unsymmetric powder pattern of the chemical shift anisotropy at the high and low temperature phase, respectively. For the deuterated compound, the correlation times of the electron spin in Cu2+ were estimated from 195Pt T1 and the activation energy for jumping between the different configurations of Jahn-Teller distortion Δ = 200 K was obtained.

Structure and dynamics of the metal site of cadmium-substituted carboxypeptidase A in solution and crystalline states and under steady-state peptide hydrolysis.

PAC spectra (perturbed angular correlation of gamma-rays) of cadmium-substituted carboxypeptidase A (CPD) show that the enzyme in solution imposes a flexible, pH- and chloride-dependent coordination structure on the metal site, in contrast to what is found in the crystalline state. A much more restricted coordination geometry occurs for the steady-state peptide intermediates of Bz-Gly-l-Phe and Bz-Gly-Gly-l-Phe in solution, suggesting that substrate binding locks the structure in a rigid conformation. The results further indicate that the peptide intermediate has a six-coordinated metal coordination geometry with an OH- ligand at the solvent site and a carbonyl oxygen at an additional ligand site. In marked contrast, conformational rigidity is not induced by the inhibitor/poor substrate Gly-L-Tyr nor by the products of high turnover substrates, Bz-Gly, Bz-Gly-Gly, and L-Phe. These results are consistent with an intact scissile peptide bond in the enzyme-substrate complex of Bz-Gly-L-Phe and Bz-Gly-Gly-L-Phe. A single nuclear quadrupole interaction (NQI) is observed for the crystalline state of the enzyme between pH 5.7 and pH 9.4. This NQI agrees with calculations based on the metal coordination geometry for cadmium in crystalline CPD derived from X-ray diffraction studies. A single broad distribution of NQIs is observed for CPD in sucrose solutions and 0.1 M NaCl at pH values below 6.5. This NQI (NQI-1') has parameters very close to those for the crystalline state. The enzyme metal site, characterized by this NQI, is converted into two new enzyme metal sites over the pH range of 6.5-8.3. The metal coordination sphere of one of these has a NQI (NQI-1) with parameters similar to those at lower pH values (NQI-1') while the other NQI (NQI-2) is characterized by markedly different NQI parameters. Angular overlap model (AOM) calculations indicate that the coordination sites giving NQI-1' and NQI-1 both have a metal-bound water molecule while the coordination site giving NQI-2 has a metal-bound hydroxide ion. PAC results at pH 8.3-10.5 indicate that in this pH range the two metal coordination geometries related to NQI-1 and NQI-2 occur in a pH independent ratio of 2:1, with the one with the water ligand being the most abundant species. The observed pH-independent equilibrium between the two different metal coordination geometries for cadmium can be explained by an equilibrium between tautomeric forms of a hydrogen bond between the Glu-270 carboxyl group and the metal-bound water (Glu-270 COO-...(HOH)M <==> Glu-270 COOH...(OH-)M) being slow on the time scale of a PAC experiment, i.e., slower than 0.5 micros. We finally suggest that NQI-1' observed at low pH reflects an enzyme species containing a metal-coordinated water molecule and the protonated carboxyl group of Glu-270.