Structural insights into the ferroxidase site of ferritins from higher eukaryotes

The first step of iron biomineralization mediated by ferritin is the oxidation at the ferroxidase active site of two ferrous ions to a diferric oxo/hydroxo species. Metal-loaded ferritin crystals obtained by soaking crystals of frog ferritin in FeSO(4) and CuSO(4) solutions followed by flash freezing provided X-ray crystal structures of the tripositive iron and bipositive copper adducts at 2.7 and 2.8 ? resolution, respectively. At variance with the already available structures, the crystal form used in this study contains 24 independent subunits in the asymmetric unit permitting comparison between them. For the first time, the diferric species at the ferroxidase site is identified in ferritins from higher eukaryotes. Anomalous difference Fourier maps for crystals (iron crystal 1) obtained after long soaking times in FeSO(4) solution invariantly showed diferric species with a Fe-Fe average distance of 3.1 ± 0.1 ?, strongly indicative of the presence of a μ-oxo/hydroxo bridge between the irons; protein ligands for each iron ion (Fe1 and Fe2) were also unequivocally identified and found to be the same in all subunits. For copper bound ferritin, dicopper(II) centers are also observed. While copper at site 1 is essentially in the same position and has the same coordination environment as Fe1, copper at site 2 is displaced toward His54, now acting as a ligand; this results in an increased intermetal distance (4.3 ± 0.4 ?). His54 coordination and longer metal-metal distances might represent peculiar features of divalent cations at the ferroxidase site. This oxidation-dependent structural information may provide key features for the mechanistic pathway in ferritins from higher eukaryotes that drive uptake of bivalent cation and release of ferric products at the catalytic site. This mechanism is supported by the X-ray picture obtained after only 1 min of soaking in FeSO(4) solutions (iron crystal 2) which reasonably contain the metal at different oxidation states. Here two different di-iron species are trapped in the active site, with intermetal distances corresponding to those of the ferric dimer in crystal 1 and of the dicopper centers and corresponding rearrangement of the His54 side chain.     

An inverted triplesalen ligand by a convergent synthesis and its influence on trinuclear FeIII 3 complexes

The inverted triplesalen ligand H 6 feld Me has been synthesized from 2,4,6-triformyl-phloroglucinol and a ketimine salen half-unit in a convergent synthesis. NMR, IR, and UV-vis spectroscopy reveal that this ligand is not in the O-protonated tautomer but in the N-protonated tautomer with substantial heteroradialene contribution. This ligand and the conventional triplesalen ligand H 6 talen t-Bu 2 have been used to synthesize the trinuclear Fe III complexes [(feld Me )(FeCl) 3 ] and [(talen t-Bu 2 )(FeCl) 3 ], respectively. The molecular structures of these complexes were obtained by single-crystal X-ray diffraction. Two trinuclear Fe III complexes of [(feld Me )(FeCl) 3 ] dimerize via two Fe-phenolate bonds, whereas due to steric hindrance no dimerization is observed for [(talen t-Bu 2 )(FeCl) 3 ]. The structural data also reveal some heteroradialene contribution in the trinuclear complexes. Whereas UV-vis and M bauer spectroscopy are not suitable to distinguish between the two complexes, FT-IR spectra show characteristic features due to the different substitution patterns of the conventional and the inverted triplesalen ligands. Another handle is provided by electrochemistry. Whereas both complexes exhibit an irreversible oxidation wave (0.94 V vs. Fc + /Fc for [(feld Me )(FeCl) 3 ] and 0.84 V vs. Fc + /Fc for [(talen t-Bu 2 )(FeCl) 3 ]), which is assigned to the oxidation of the central backbone, higher potential oxidations are reversible for [(talen t-Bu 2 )(FeCl) 3 ]) but irreversible for [(feld Me )(FeCl) 3 ]. This is attributed to the reversible oxidation of the terminal phenolates in the di-tert-butyl substituted [(talen t-Bu 2 )(FeCl) 3 ] in contrast to the mono-methyl-substituted phenolates in [(feld Me )(FeCl) 3 ]. The magnetic properties of [(talen t-Bu 2 )(FeCl) 3 ] reveal a very small ferromagnetic coupling with significant zero-field splitting of the FeⅢ S = 5/2 ions. In contrast, the dimerization of two trinuclear complexes in [(feld Me )(FeCl) 3 ] results in antiferromagnetic interactions between the two phenolate-bridged Fe III ions, which mask the intra-trinuclear interactions transmitted by the central phloroglucinol backbone.     

Quasi-equilibrium states of copolymers that simultaneously crystallize and interchange structural units

A quasi-equilibrium model of a step-growth copolymer that crystallizes and interchanges structural units simultaneously is presented. Equilibrium values of counit-sequence-length distribution in the amorphous phase and degree of crystallinity are calculated in a system for which crystallite thickness is determined by nucleation. When applied to calculations, the model predicts that the counits of the amorphous phase will tend to be ordered in blocks. The crystalline content of the whole polymer and the extent of ordering counits along the chain decreases with increasing temperature. The model predicts temperature ranges for some whole copolymers in which one of two similarly stable, disparate combinations of crystallinity and sequence-length distribution may prevail.     

Experimental and theoretical investigation of the photoionization of hydrogen cyanide

Cryogenic matrices containing molecular aggregates of methyl nitrite or methanol have been irradiated with a cw CO₂ laser. At the irradiation frequency hole burning was observed by means of a grating infrared spectrophotometer. The results suggest that vibrational absorption lineshapes of aggregates in cryogenic matrices are dominated by inhomogeneous line-broadening.     

Validity and Failure of the Cauchy-Born Hypothesis in a Two-Dimensional Mass-Spring Lattice

Summary. The Cauchy-Born rule postulates that when a monatomic crystal is subjected to a small linear displacement of its boundary, then all atoms will follow this displacement. In the absence of previous mathematical results, we study the validity of this rule in the model case of a 2D cubic lattice interacting via harmonic springs between nearest and diagonal neighbours. Our main result is that for favourable values of the spring constants and spring equilibrium lengths, the CB rule is actually a theorem. Simple counterexamples show that for unfavourable spring parameters or large displacements the CB rule fails. Moreover the resulting overestimation of the lattice energy per unit volume by the CB rule cannot be cured by convexification (let alone quasiconvexification) of the CB energy. The main tool in our proof is a novel notion of lattice polyconvexity which allows us to overcome the difficulty that the elastic energy as a function of atomic positions can never be convex, due to frame-indifference.     

A Proof of Crystallization in Two Dimensions

Many materials have a crystalline phase at low temperatures. The simplest example where this fundamental phenomenon can be studied are pair interaction energies of the type where y(x) ∈ℝ² is the position of particle x and V(r) ∈ ℝ is the pair-interaction energy of two particles which are placed at distance r. Due to the Mermin-Wagner theorem it can't be expected that at finite temperature this system exhibits long-range ordering. We focus on the zero temperature case and show rigorously that under suitable assumptions on the potential V which are compatible with the growth behavior of the Lennard-Jones potential the ground state energy per particle converges to an explicit constant E_*: where E_* ∈ ℝ is the minimum of a simple function on [0,∞). Furthermore, if suitable Dirichlet- or periodic boundary conditions are used, then the minimizers form a triangular lattice. To the best knowledge of the author this is the first result in the literature where periodicity of ground states is established for a physically relevant model which is invariant under the Euclidean symmetry group consisting of rotations and translations.     

The effect of prior crystallinity on the flow-induced crystallization of poly(ethylene oxide)

After flowing in a dilatometer bulb for a small fraction of the duration of the transformation, a relaxed melt of poly(ethylene oxide) (M?_n = (5.9 ± 0.1) × 10³) showed marked increases in isothermal crystallization rate. The extent of increase was greater when flow was imposed at modestly later stages rather than at the earliest stage of a crystallization. Kinetic parameters for the flow-induced crystallizations were obtained via modification of the conventional mathematical treatment of the kinetics of phase change, thereby allowing the analytical resolution of the overall process into flowinduced and quiescent components. Determination of the flow-induced crystallization parameters required independent determination of the kinetic parameters for quiescent crystallizations at that temperature. The Avrami exponents n_f which characterized the flow-induced portions of the crystallizations were larger for those instances in which flow was imposed at the more advanced stages of the crystallizations, thus indicating a transition in crystallization mechanism. It is suggested that prior crystallinity present at the time of flow contributed to the crystallization by serving as a source of nucleation sites. However, in light of the experimental procedure employed, values of n_f approximating 4 that were obtained are not susceptible to mechanistic interpretations now extant.     

Spectroscopic definition of the ferroxidase site in M ferritin: comparison of binuclear substrate vs cofactor active sites

Maxi ferritins, 24 subunit protein nanocages, are essential in humans, plants, bacteria, and other animals for the concentration and storage of iron as hydrated ferric oxide, while minimizing free radical generation or use by pathogens. Formation of the precursors to these ferric oxides is catalyzed at a nonheme biferrous substrate site, which has some parallels with the cofactor sites in other biferrous enzymes. A combination of circular dichroism (CD), magnetic circular dichroism (MCD), and variable-temperature, variable-field MCD (VTVH MCD) has been used to probe Fe(II) binding to the substrate active site in frog M ferritin. These data determined that the active site within each subunit consists of two inequivalent five-coordinate (5C) ferrous centers that are weakly antiferromagnetically coupled, consistent with a mu-1,3 carboxylate bridge. The active site ligand set is unusual and likely includes a terminal water bound to each Fe(II) center. The Fe(II) ions bind to the active sites in a concerted manner, and cooperativity among the sites in each subunit is observed, potentially providing a mechanism for the control of ferritin iron loading. Differences in geometric and electronic structure--including a weak ligand field, availability of two water ligands at the biferrous substrate site, and the single carboxylate bridge in ferritin--coincide with the divergent reaction pathways observed between this substrate site and the previously studied cofactor active sites.     

On the behavior of dissipative systems in contact with a heat bath: Application to Andrade creep

We develop a theory of statistical mechanics for dissipative systems governed by equations of evolution that assigns probabilities to individual trajectories of the system. The theory is made mathematically rigorous and leads to precise predictions regarding the behavior of dissipative systems at finite temperature. Such predictions include the effect of temperature on yield phenomena and rheological time exponents. The particular case of an ensemble of dislocations moving in a slip plane through a random array of obstacles is studied numerically in detail. The numerical results bear out the analytical predictions regarding the mean response of the system, which exhibits Andrade creep.