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The biological N2-fixation reaction is catalyzed by the enzyme nitrogenase. The metal cluster active site of this enzyme, the iron-molybdenum cofactor (FeMoco), can be studied either while bound within the MoFe protein component of nitrogenase or after it has been extracted into N-methylformamide. The two species are similar but not identical. For example, the addition of thiophenol or selenophenol to isolated FeMoco causes its rather broad S = 3/2 electron paramagnetic resonance signal to sharpen and more closely approach the signal exhibited by protein-bound FeMoco. The nature of this thiol/selenol binding site has been investigated by using Se-K edge extended x-ray absorption fine structure (EXAFS) to study selenophenol ligated to FeMoco, and the results are reported here. EXAFS data analysis at the ligand Se-K edge was performed with a set of software, GNXAS, that provides for direct calculation of the theoretical EXAFS signals and least-squares fits to the experimental data. Data analysis results show definitively that the selenol (and by inference thiol) binds to Fe at a distance of 2.4 A. In contrast, unacceptable fits are obtained with either Mo or S as the liganded atom (instead of Fe). These results provide quantitative details about an exchangeable thiol/selenol binding site on FeMoco in its isolated, solution state and establish an Fe atom as the site of this reaction. Furthermore, the utility of ligand-based EXAFS as a probe of coordination in polynuclear metal clusters is demonstrated.

X-ray absorption spectroscopy (XAS) of Fe(2+) in Fe(2)SiO(4) liquid at 1575 kelvin and 10(-4) gigapascal (1 bar) shows that the Fe(2+) -O bond length is 1.98 +/- 0.02 angstroms compared with approximately 2.22 angstroms in crystalline Fe(2)SiO(4) (fayalite) at the melting point (1478 kelvin), which indicates a decrease in average Fe(2+) coordination number from six in fayalite to four in the liquid. Anharmonicity in the liquid was accounted for using a data analysis procedure. This reduction in coordination number is similar to that observed on the melting of certain ionic salts. These results are used to develop a model of the medium-range structural environment of Fe(2+) in olivine-composition melts, which helps explain some of the properties of Fe(2)SiO(4) liquid, including density, viscosity, and the partitioning of iron and nickel between silicate melts and crystalline olivines. Some of the implications of this model for silicate melts in the Earth's crust and mantle are discussed.

Copper K-edge x-ray absorption data indicate that an axial oxygen-centered lattice instability accompanying the 93 K superconducting transition in YBa(2)Cu(3)O(7) is of a pseudo-(anti)ferroelectric type, in that it appears to involve the softening of a double potential well into a structure in which the difference between the two copper-oxygen distances and the barrier height have both decreased. This softer structure is present only at temperatures within a fluctuation region around the transition. A similar process involving the analogous axial oxygen atom also accompanies the superconducting transition in T1Ba(2)Ca(3)Cu(4)O(11), where the superconducting transition temperature T(c) is ~120 K. The mean square relative displacement of this oxygen atom in YBa(2)Cu(3)O(7) is also specifically affected by a reduction in the oxygen content and by the substitution of cobalt for copper, providing further evidence for the sensitivity of the displacement to additional factors that also influence the superconductivity. On the basis of the implied coupling of this ionic motion to the superconductivity, a scenario for high-temperature superconductivity is presented in which both phonon and electronic (charge transfer) channels are synergistically involved.

The reaction of the iron-molybdenum cofactor with thiolate and the redox behavior of the iron-molybdenum cofactor-thiolate complex have been studied by 19F NMR using p-CF3C6H4S- as the reporter ligand. These experiments give results different from those produced by other methods which have been performed near 4 K rather than at ambient temperature. Specifically, these data show that the iron-molybdenum cofactor-thiolate complex is not the product of an irreversible reaction. Rather, the complex is in dynamic equilibrium with the free iron-molybdenum cofactor and free thiolate. Models of the reactions of nitrogenase may need to take this temperature-dependent difference into account because the lability of the iron-molybdenum thiolate bond means its making and breaking could be involved in substrate binding or reduction. The 19F NMR results reported here also show that the S = 3/2 state of the iron-molybdenum cofactor-thiolate complex can be easily and reversibly oxidized by one electron. However, electron exchange between the oxidized and reduced states of the complex is quite slow at approximately 1 mM. Based on low temperature spectroscopic studies, the oxidized iron-molybdenum cofactor-thiolate complex was expected to be diamagnetic. Isotropically shifted NMR spectra of the oxidized cofactor samples at 240-320 K, however, indicate at least partial population of a paramagnetic state, possibly with S = 1.

Changes in the copper K-edge x-ray absorption spectrum of YBa(2)Cu(3)O(7) across the critical temperature indicate that, accompanying the superconducting transition, the mean square relative displacement of some fraction of the Cu2-O4 bonds becomes smaller or more harmonic (or both), that there may be a slight increase in the associated Cul-O4 distance, and that electronic states involving these atom pairs become more atomic-like. If there is an association between the superconductivity and this lattice instability, then the bridging axial oxygen is of central importnce in determining the high tranition temperature of YBa(2)Cu(3)O(7). Because this structural perturbation will affect the dynamic polrizability of the copper oxygen sublattice, it is consistent with an excitonic pairing mhanism in these materials.

19F NMR and x-ray absorption experiments have been performed with both the isolated FeMo cofactor and the MoFe protein of nitrogenase in search of direct evidence for substrate or inhibitor binding. Using 19F NMR as a probe and p-CF3C6H4S- as the receptor ligand, the data show that the nitrogenase inhibitors CN- and CH3NC bind to the isolated FeMo cofactor-RFS- complex in N-methylformamide with a finite formation constant. Their binding increases the electronic relaxation time of the complex and increases the life-time of the FeMo cofactor-p-CF3C6H4S- bond, Parallel molybdenum K edge and extended x-ray absorption fine structure experiments show that CH3NC does not bind to molybdenum. Although CO and N3- both relieve CN- and CH3NC inhibition of electron flow through nitrogenase, unlike the latter, they do not appear to bind to isolated FeMo cofactor. In experiments with the dithionite-reduced MoFe protein, we did not detect any changes in the molybdenum K edge or extended x-ray absorption fine structure spectra upon addition of CO, N2, C2H2, NaCN, CH3NC, or azide demonstrating that either these substrates and inhibitors do not bind to molybdenum or that the FeMo cofactor site of nitrogenase is inaccessible to substrate binding except under turnover conditions.