Resonance Raman and infrared spectroscopic studies of high-output forms of human soluble guanylyl cyclase

The allosteric regulator BAY-41-2272 converts the CO adduct of soluble guanylyl cyclase (CO-sGC) enzyme from a low- to high-output form, with respect to production of cGMP. Resonance Raman (RR) and Fourier Transform Infrared (FTIR) spectroscopic techniques are used to show that the CO-sGC exists as major and minor conformers, both having nu(Fe-CO) and nu(C-O) modes characteristic of 6-coordinate species. It is further shown that addition of BAY-41-2272 to the CO adduct induces the transition of some fraction of the initial CO-heme adducts into two new CO-heme complexes, the fractional conversion being dependent on the temperature. One new complex displays vibrational modes characteristic of pentacoordinated CO-adduct, and its formation is not affected by temperature. The second complex, although slightly different from the original CO-adducts, is hexacoordinated, and its formation is facilitated by temperature. The production of substantial amounts of the 5-coordinate CO adduct upon addition of BAY-41-2272, reveals the fact that several out-of-plane heme deformation modes are simultaneously activated, an observation similar to that realized upon NO activation. While the precise nature of these modes will require elucidation by isotopic labeling experiments, by analogy with earlier studies of other heme proteins, several bands associated with modes attributable to peripheral substituent deformations and methine carbon movements are implicated. The documented formation of two new forms upon addition of Bay-41-2272 (a 5-coordinate and a new 6-coordinate form) is discussed with respect to the implications for enzyme activation.     

Investigation of solvent effects for the Claisen rearrangement of chorismate to prephenate: mechanistic interpretation via near attack conformations

Solvent effects on the rate of the Claisen rearrangement of chorismate to prephenate have been examined in water and methanol. The preequilibrium free-energy differences between diaxial and diequatorial conformers of chorismate, which had previously been implicated as the sole basis for the observed 100-fold rate increase in water over methanol, have been reframed using the near attack conformation (NAC) concept of Bruice and co-workers. Using a combined QM/MM Monte Carlo/free-energy perturbation (MC/FEP) method, 82%, 57%, and 1% of chorismate conformers were found to be NAC structures (NACs) in water, methanol, and the gas phase, respectively. As a consequence, the conversion of non-NACs to NACs provides no free-energy contributions to the overall relative reaction rates in water versus methanol. Free-energy perturbation calculations yielded differences in free energies of activation for the two polar protic solvents and the gas phase. The rate enhancement in water over the gas phase arises from preferential hydration of the transition state (TS) relative to the reactants via increased hydrogen bonding and long-range electrostatic interactions, which accompany bringing the two negatively charged carboxylates into closer proximity. More specifically, there is an increase of 1.3 and 0.6 hydrogen bonds to the carboxylate groups and the ether oxygen, respectively, in going from the reactant to the TS in water. In methanol, the corresponding changes in hydrogen bonding with first shell solvent molecules are small; the rate enhancement arises primarily from the enhanced long-range interactions with solvent molecules. Thus, the reaction occurs faster in water than in methanol due to greater stabilization of the TS in water by specific interactions with first shell solvent molecules.     

Calculation of optimum geometries and force fields by the CNDO/force method

CNDO/Force calculations have been performed on a series of molecules, H₂CO, F₂CO, CF₄, CHF₃, CH₂F₂ and CH₃F. The optimum geometries and force fields are reported. It is found that the method can successfully predict the geometries of polyatomic molecules. The bending force constants and interaction force constants are, in general, comparable with experimental values both with respect to sign and magnitude. The stretching force constants have higher values than the experimental force constants. However, the trend in stretching force constants of a series of molecules is comparable with that of the corresponding experimental values.     

Photoswitchable multivalent sugar ligands: synthesis, isomerization, and lectin binding studies of azobenzene-glycopyranoside derivatives

Coating of azobenzene chromophore with multivalent sugar ligands has been accomplished. Such sugar coating allows the study of the isomerization properties of this chromophore in aqueous solutions. The predominantly cis-isomer-containing photostationary state (PS) mixture of these azobenzene derivatives is found to be stable for hours. The rate constants for their isomerization, as well as the Arrhenius activation energies, are determined experimentally. An assessment of the lectin binding properties of the lactoside bearing isomeric azobenzene derivatives, by isothermal calorimetric methods, reveals the existence of an unusual cooperativity in their binding to lectin peanut agglutinin. Thermodynamic parameters evaluated for the trans and the PS mixture are discussed, in detail, for the lactoside bearing bivalent azobenzene derivative.     

Stabilization of methyl anions by first-row substituents. The superiority of diffuse function-augmented basis sets for anion calculations

The entire set of methyl anions, XCH₂^?, substituted by first-row substituents, Li, BeH, BH₂, CH₃, NH₂, OH, and F, was examined at various ab initio levels. Diffuse orbital-augmented basis sets, such as 4?31+G and 6?31+G *, are needed to describe the energies of these anions adequately. Estimates of proton affinities are further improved by second-order Møller–Plesset (MP 2) electron correlation corrections, but relative energies are less affected. The methyl group in the ethyl anion is destabilizing, the amino substituent is borderline, but all other groups are stabilizing. Very large π effects are exhibited by BH₂ and BeH groups; inductive stabilization by the electronegative F and OH groups is less effective. Lithium also is stabilizing, but the best singlet geometry of CH₂Li^? is not planar. A planar CH₂Li^? triplet with a π¹ configuration may be lower in energy.     

Carbodications. I. The structures and energies of C4H isomers

At high levels of ab initio theory (6-31G*//4-31G), the most stable C₄H isomer is indicated to be the nonplanar cyclobutadiene dication ( 1a ); the planar form, 1b , is indicated to be 7.5 kcal/mol less stable. The second most stable C₄H isomer, the methylenecyclopropene dication, is indicated to prefer the perpendicular ( 2a ) over the planar ( 2b ) arrangement by 7 kcal/mol. The “anti van't Hoff” cyclo-(HB)₂C?CH₂ system ( 4 ), isoelectronic with 2 , also prefers the perpendicular conformation ( 4a ), and retains the C?C double bond. The linear butatriene dication ( 3 ) is the least stable C₄H species investigated. The perpendicular (D_2d) arrangement ( 3a ), permitting double allyl cationlike conjugation, is preferred over the planar D_2h form ( 3b ) by 26 kcal/mol. The heat of formation of the most stable form of C₄H, 1a , is estimated to be 623–640 kcal/mol. This species should be thermodynamically stable toward dissociation into smaller charged fragments.     

Diastereo- and Enantioselective Synthesis of syn-alpha-Vinylchlorohydrins and cis-Vinylepoxides

A new method to generate chiral syn-vinylchlorohydrins and cis-vinyloxiranes is reported. Reaction of (alpha-haloallyl)lithiums with methoxy-9-BBN or Ipc(2)BOMe followed by treatment with BF(3).OEt(2) leads to (Z)-(gamma-haloallyl)boranes which react with aldehydes to yield cis-vinylepoxides (de >/= 90%) upon oxidative workup. Alternatively, addition of ethanolamine to the allylboration product yields syn-alpha-halohydrins (de >/= 90%) that are also easily cyclized to cis-vinylepoxides. Extension of this protocol using [(Z)-gamma-chloroallyl]BIpc(2) leads to chiral syn-alpha-chlorohydrins and cis-vinylepoxides in high de (>/=90%) and ee (90-99%). Enantioselectivity of reactions of chiral (Z)-(gamma-chloroallyl)boranes with aldehydes are more sensitive to reaction conditions than enantioselectivity of reactions of other alpha-or gamma-substituted allylboranes. The effects of proportion of BF(3).OEt(2) and the relative efficacies of LiNR(2) bases on diastereo- and enantioselectivity of the chloroallylation are reported.     

Characterization of non-linear adsorption properties of dextran-based polyelectrolyte displacers in ion-exchange systems

Experimental studies were carried out on the non-linear adsorption properties of dextran-based polyelectrolytes in anion- and cation-exchange chromatographic systems. By monitoring both the induced salt gradients and sequential breakthrough fronts, parameters were determined for use in a Steric Mass Action (SMA) model of non-linear ion-exchange chromatography. These parameters include: total ion capacity of the columns, characteristic charge, steric factor, equilibrium constant, and maximum adsorptive capacity for each of the polyelectrolytes. In addition the number of functional groups were determined by elemental analysis. The values of the SMA parameters were found to be independent of salt and polyelectrolyte bulk phase compositions. Parameters were also determined for a variety of proteins. Experimental isotherms for the polyelectrolytes and proteins were compared with those simulated by the SMA model. Finally, the implications of polyelectrolyte adsorption properties with respect to their ability to act as efficient displacers in ion-exchange displacement systems are discussed.     

High performance dye-sensitized solar cell based on 2-mercaptobenzimidazole doped poly(vinylidinefluoride-co-hexafluoropropylene) based polymer electrolyte

In this work, the influence of 2-mercaptobenzimidazole (2-MCBI) on poly(vinylidinefluoride-co-hexafluoropropylene)/KI/I₂ (PVDF-HFP/KI/I₂) polymer electrolytes were studied. The pure and different weight percentage ratios (20, 30, 40 and 50%) of 2-MCBI doped PVDF-HFP/KI/I₂ electrolytes were prepared by a solution casting technique. The as-prepared polymer electrolyte films were characterized using various techniques such as Fourier transform infrared (FT-IR) spectroscopy, differential scanning calorimetry (DSC), X-ray diffractometer (XRD), alternating current (AC)-impedance analysis. The addition of 2-MCBI with pure PVDF-HFP/KI/I₂ was found to increase the ionic conductivity of electrolyte. Among the various additions, 30 wt% 2-MCBI doped PVDF-HFP/KI/I₂ showed the highest room temperature ionic conductivity values than the others. The dye-sensitized solar cell (DSSC) fabricated using this optimized polymer electrolyte achieved a high power conversion efficiency of 4.40% than the pure PVDF-HFP/KI/I₂ (1.74%) at similar experimental conditions. Thus, the 2-MCBI doped polymer electrolyte has proven to be an effective substitute to the liquid electrolyte in DSSCs.