Extended X-ray absorption fine structure and nuclear resonance vibrational spectroscopy reveal that NifB-co, a FeMo-co precursor, comprises a 6Fe core with an interstitial light atom

NifB-co, an Fe-S cluster produced by the enzyme NifB, is an intermediate on the biosynthetic pathway to the iron molybdenum cofactor (FeMo-co) of nitrogenase. We have used Fe K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy together with (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to probe the structure of NifB-co while bound to the NifX protein from Azotobacter vinelandii. The spectra have been interpreted in part by comparison with data for the completed FeMo-co attached to the NafY carrier protein: the NafY:FeMo-co complex. EXAFS analysis of the NifX:NifB-co complex yields an average Fe-S distance of 2.26 A and average Fe-Fe distances of 2.66 and 3.74 A. Search profile analyses reveal the presence of a single Fe-X (X = C, N, or O) interaction at 2.04 A, compared to a 2.00 A Fe-X interaction found in the NafY:FeMo-co EXAFS. This suggests that the interstitial light atom (X) proposed to be present in FeMo-co has already inserted at the NifB-co stage of biosynthesis. The NRVS exhibits strong bands from Fe-S stretching modes peaking around 270, 315, 385, and 408 cm(-1). Additional intensity at approximately 185-200 cm(-1) is interpreted as a set of cluster "breathing" modes similar to those seen for the FeMo-cofactor. The strength and location of these modes also suggest that the FeMo-co interstitial light atom seen in the crystal structure is already in place in NifB-co. Both the EXAFS and NRVS data for NifX:NifB-co are best simulated using a Fe 6S 9X trigonal prism structure analogous to the 6Fe core of FeMo-co, although a 7Fe structure made by capping one trigonal 3S terminus with Fe cannot be ruled out. The results are consistent with the conclusion that the interstitial light atom is already present at an early stage in FeMo-co biosynthesis prior to the incorporation of Mo and R-homocitrate.     

Fourier transform infrared spectroscopic study of water-in-supercritical CO2 microemulsion as a function of water content

Fourier transform infrared (FT-IR) spectrum of water-in-supercritical CO(2) microemulsion was measured at 60 degrees C and 30.0 MPa over a wide range of water/CO(2) ratio from 0.0 to 1.2 wt % to study the distribution of water into CO(2), interfacial area around surfactant headgroup, and core water pool. The microemulsion was stabilized by sodium bis(1H,1H,2H, 2H-heptadecafluorodecyl)-2-sulfosuccinate [8FS(EO)(2)] equimolarly mixed with sodium 1-oxo-1-[4-(tridecafluorohexyl)phenyl]-2-hexanesulfonate [FC6HC4] or with poly(ethylene glycol) 2,6,8-trimethyl-4-nonyl ether [TMN-6]. The signal area of the O-H stretching band of water suggested that the number of water molecules in the microemulsion increases linearly with the water/CO(2) ratio, except for a slow initial increase below 0.4 wt % due to a part of water dissolved in CO(2). The amount of water in CO(2) was evaluated by decomposing the bending band of water into two components, one at lower frequency ascribed to water in CO(2) and the other at higher frequency to water in the microemulsion. The decomposition confirmed that CO(2) is saturated with water at the water content of 0.4 wt %. It was also revealed, from the symmetric SO stretching frequency of the surfactant, that the sulfonate headgroup is completely hydrated at the water/CO(2) ratio of 0.4-0.5 wt %. The results demonstrated that water is introduced preferentially into CO(2) and the interfacial area at small water content, and then is loaded into the micelle core after the saturation of CO(2) with water and the full hydration of the surfactant headgroup.     

Characterization of the Fe site in iron-sulfur cluster-free hydrogenase (Hmd) and of a model compound via nuclear resonance vibrational spectroscopy (NRVS)

We have used (57)Fe nuclear resonance vibrational spectroscopy (NRVS) to study the iron site in the iron-sulfur cluster-free hydrogenase Hmd from the methanogenic archaeon Methanothermobacter marburgensis. The spectra have been interpreted by comparison with a cis-(CO)2-ligated Fe model compound, Fe(S2C2H4)(CO)2(PMe3)2, as well as by normal mode simulations of plausible active site structures. For this model complex, normal mode analyses both from an optimized Urey-Bradley force field and from complementary density functional theory (DFT) calculations produced consistent results. For Hmd, previous IR spectroscopic studies found strong CO stretching modes at 1944 and 2011 cm(-1), interpreted as evidence for cis-Fe(CO)2 ligation. The NRVS data provide further insight into the dynamics of the Fe site, revealing Fe-CO stretch and Fe-CO bend modes at 494, 562, 590, and 648 cm(-1), consistent with the proposed cis-Fe(CO)2 ligation. The NRVS also reveals a band assigned to Fe-S stretching motion at approximately 311 cm(-1) and another reproducible feature at approximately 380 cm(-1). The (57)Fe partial vibrational densities of states (PVDOS) for Hmd can be reasonably well simulated by a normal mode analysis based on a Urey-Bradley force field for a five-coordinate cis-(CO)2-ligated Fe site with additional cysteine, water, and pyridone cofactor ligands. A "truncated" model without a water ligand can also be used to match the NRVS data. A final interpretation of the Hmd NRVS data, including DFT analysis, awaits a three-dimensional structure for the active site.     

Observations of the near-wall accumulation of suspended particles due to shear and electroosmotic flow in opposite directions

On the basis of previous studies, the particles in a dilute (volume fractions φ_∞ < 4 × 10^–3) suspension in combined Poiseuille and electroosmotic “counterflow” at flow Reynolds numbers Re ≤ 1 accumulate, then assemble into structures called “bands,” within ∼6 μm of the channel wall. The experimental studies presented here use a small fraction of tracer particles labeled with a different fluorophore from the majority “bulk” particles to visualize the dynamics of individual particles in a φ_∞ = 1.7 × 10^–3 suspension. The results at two different near-wall shear rates and three electric field magnitudes E show that the near-wall particles are concentrated about 150-fold when the bands start to form, and are then concentrated about 200-fold to a maximum near-wall volume fraction of ∼0.34. The growth in the near-wall particles during this accumulation stage appears to be exponential. This near-wall particle accumulation is presumably driven by a wall-normal “lift” force. The observations of how the particles accumulate near the wall are compared with recent analyses that predict that suspended particles subject to shear flow and a dc electric field at small particle Reynolds numbers experience such a lift force. A simple model that assumes that the particles are subject to this lift force and Stokes drag suggests that the force driving particles toward the wall, of O(10^–17 N), is consistent with the time scales for particle accumulation observed in the experiments.     

Total synthesis and absolute stereochemistry of (+)-batzellaside B and its C8-epimer, a new class of piperidine alkaloids from the sponge Batzella sp.

The first total synthesis of (+)-batzellaside B and its C8-epimer was completed from a known l-arabinose-derived tribenzyl ether in 22 steps with overall yields of 3.9% and 5.4%, respectively. The absolute configuration of (+)-batzellaside B was unambiguously determined to be 1S,3S,4S,5R,8S by the Mosher analysis of a synthetic intermediate prepared through a separate route.     

Influence of the chelate effect on the electronic structure of one-electron oxidized group 10 metal(II)-(disalicylidene)diamine complexes

The neutral and one-electron oxidized group 10 metal, Ni(II), Pd(II) and Pt(II), six-membered chelate Salpn (Salpn = N,N'-bis(3,5-di-tert-butylsalicylidene)-1,3-propanediamine) complexes have been investigated and compared to the five-membered chelate Salen (N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-ethanediamine) and Salcn (N,N'-bis(3,5-di-tert-butylsalicylidene)-1,2-cyclohexane-(1R,2R)-diamine) complexes. Reaction of the Salpn complexes with 1 equivalent of AgSbF(6) affords the oxidized complexes which exist as ligand radical species in solution and in the solid state. The solid state structures of the oxidized complexes have been determined by X-ray crystal structure analysis. While the Ni and Pt analogues exhibit an essentially symmetric coordination sphere contraction upon oxidation, the oxidized Pd derivative exhibits an asymmetric metal binding environment demonstrating at least partial ligand radical localization. In comparison to the oxidized Salen and Salcn complexes, the propyl backbone of the Salpn complexes leads to a larger deviation from a planar geometry in the solid state. The electronic structure of the oxidized Salpn complexes was further probed by UV-vis-NIR measurements, electrochemistry, EPR spectroscopy, and theoretical calculations. The intense NIR band for the one-electron oxidized Salpn complexes shifts to lower energy in comparison to the 5-membered chelate analogues, which is attributed to lower metal d(xz) character in the β-LUMO for the Salpn series. The reactivity of the one-electron oxidized Salpn complexes with exogenous ligands was also studied. In the presence of pyridine, the oxidized Ni analogue exhibits a shift in the locus of oxidation to a Ni(III) species. The oxidized PtSalpn complex rapidly decomposes in the presence of pyridine, even at low temperature. Interestingly, electronic and EPR spectroscopy suggests that the addition of pyridine to the oxidized Pd analogue results in initial dissociation of the phenoxyl radical ligand, likely due to the increased flexibility of the propyl backbone.     

Preparation of Hemispherical Polymer Particles by Cleavage of a Janus Poly(methyl Methacrylate) /Polystyrene Composite Particle

Micrometer-sized, hemispherical particles were successfully prepared as a result of the cleavage of Janus PMMA/PS composite particles by dispersion into acetone/water (9/1-10/0 v/v) media or a THF/water (8/2 v/v) medium. The spherical composite particles having a Janus structure were prepared by the slow evaporation of toluene from homogeneous PMMA/PS/toluene droplets dispersed in an aqueous medium in advance. It was clarified that the difference in affinity between PMMA and PS phases with respect to the media caused the cleavage of the composite particles. This method is expected to be a novel approach to the preparation of nonspherical polymer particles.     

Reverse enantioselectivity in the lipase-catalyzed desymmetrization of prochiral 2-carbamoylmethyl-1,3-propanediol derivatives

Enantioselective acetylation of 2-carbamoylmethyl-1,3-propanediol derivatives was catalyzed effectively by lipase PS to give monoacetates with high enantioselectivity: The enantioselectivity depended on the 2-carbamoylmethyl groups. The reaction of N-monoalkylcarbamoylmethyl-1,3-propanediol afforded the monoacetate with the (S)-configuration, whereas N,N-dialkylcarbamoylmethyl-1,3-propanediol gave the monoacetate with the (R)-configuration.     

Energy calibration issues in nuclear resonant vibrational spectroscopy: observing small spectral shifts and making fast calibrations

The conventional energy calibration for nuclear resonant vibrational spectroscopy (NRVS) is usually long. Meanwhile, taking NRVS samples out of the cryostat increases the chance of sample damage, which makes it impossible to carry out an energy calibration during one NRVS measurement. In this study, by manipulating the 14.4 keV beam through the main measurement chamber without moving out the NRVS sample, two alternative calibration procedures have been proposed and established: (i) an in situ calibration procedure, which measures the main NRVS sample at stage A and the calibration sample at stage B simultaneously, and calibrates the energies for observing extremely small spectral shifts; for example, the 0.3 meV energy shift between the 100%‐⁵⁷Fe‐enriched [Fe₄S₄Cl₄]⁼ and 10%‐⁵⁷Fe and 90%‐⁵⁴Fe labeled [Fe₄S₄Cl₄]⁼ has been well resolved; (ii) a quick‐switching energy calibration procedure, which reduces each calibration time from 3–4 h to about 30 min. Although the quick‐switching calibration is not in situ, it is suitable for normal NRVS measurements.     

Time-domain interferometry experiments using multi-line nuclear absorbers

Langmuir-Blodgett (LB) films of octadecanoyl hydroxamic acid (C18N) complexed with Fe^3?+? ions have been prepared at various subphase pH values. The LB films consisting of different number of layers were investigated by ⁵⁷Fe conversion electron M?ssbauer spectroscopy (CEM) at room temperature. The CEM detector contained a piece of α-iron, enriched with ⁵⁷Fe, using as an internal standard. The M?ssbauer pattern of the C18N/Fe LB films is a doublet with parameters δ = 0.35?mm/s and Δ = 0.74?mm/s. A gradual increase of the relative occurrence of the doublet compared to the sextet of the internal standard was observed with the increasing number of layers, indicating the nearly uniform distribution of Fe among the LB layers.