Theoretical investigations into the intermediacy of chlorinated vinylcobalamins in the reductive dehalogenation of chlorinated ethylenes

The reductive dehalogenation of perchloroethylene and trichloroethylene by vitamin B(12) produces approximately 95% (Z)-dichloroethylene (DCE) and small amounts of (E)-DCE and 1,1-DCE, which are further reduced to ethylene and ethane. Chloroacetylene and acetylene have been detected as intermediates, but not dichloroacetylene. Organocobalamins (RCbls) have been proposed to be intermediates in this process. Density functional theory based approaches were employed to investigate the properties of chlorinated vinylcobalamins and chlorinated vinyl radicals. They reveal that all vinyl radicals studied have reduction potentials more positive (E degrees >or= -0.49) than that of the Co(II)/Co(I) couple of B(12) (E degrees = -0.61 V), indicating that any (chlorinated) vinyl radicals formed in the reductive dehalogenation process should be reduced to the corresponding anions by cob(I)alamin in competition with their combination with Co(II) to yield the corresponding vinylcobalamins. The computed Co-C homolytic bond dissociation enthalpies (BDEs) of the latter complexes range from 33.4 to 45.8 kcal/mol. The substituent effects on the BDEs are affected by the stabilities of the vinyl radicals as well as steric interactions between (Z)-chloro substituents and the corrin ring. The calculated E degrees values of the cobalamin models were within approximately 200 mV of one another since electron attachment is to a corrin ring pi-orbital, whose energy is relatively unaffected by chloride substitution of the vinyl ligand, and all were >500 mV more negative than that of the Co(II)/Co(I) couple of B(12). Reduction of the base-off forms of vinyl- and chlorovinylcobalamin models also involves the corrin pi* orbital, but reduction of the base-off dichlorovinyl- and trichlorovinylcobalamin models occurs with electron attachment to the sigma(Co)(-)(C*) orbital, yielding calculated E degrees values more positive than that of the calculated Co(II)/Co(I) couple of B(12). Thus, cob(I)alamin is expected to reduce these base-off vinyl-Cbls. Heterolytic cleavage of the Co-C bonds is much more favorable than homolysis (>21 kcal/mol) and is significantly more exergonic when coupled to chloride elimination.     

COMPARISON OF THE EFFECTS OF ULTRAVIOLET RADIATION ON THE INTRACELLULAR CATALASE OF AEROBIC AND ANAEROBIC YEAST

Abstract— Suspensioris of aerobic and anaerobic yeast were subjected to ultraviolet radiation (principally 254 mµ ) under closely comparable experimental conditions, and changes in the level and in the temperature dependence of their catalase activity were determined. Qualita tively, the effects of U.V. on the enzyme of the anaerobic cells were similar to those on that of the aerobic cells. The effect of U.V. on the anaerobic catalase differed from that on the aerobic enzyme in the following respects: I, a considerably greater dose of U.V. was necessary in order to attain the maximum activity and the minimum activation energy of the enzyme-substrate system; 2, a far greater dose was required before appreciable photoinactivation of the maxi mally active enzyme occurred; 3, photoinactivation proceeded at less than one-half the rate; 4, the u.v.-induced increase in the catalase activity of the suspension was virtually complete before appreciable reduction in activation energy occured. The first three of these differences were interpreted in terms of a model, which pictures the anaerohic catalase as being tightly bound to an intracellular chromophore group.     

The dependence of the branching ratio in the F+HD reaction on the initial rotational state of HD

Both classical trajectory and quantal scattering calculations indicate that the branching ratio in the F+HD reaction varies considerably with the initial rotational state of HD. Information theory argues that this variation must be reflected in the distribution of the reaction products. Hence, given the (normalized) product distribution for each reaction path one should be able to predict the dependence of the branching ratio on the state of the reagents. The trajectory computations of Muckerman are used to illustrate the procedure. First the dynamic constraint is identified and then the reaction probability matrix is constructed. The determination (“synthesis”) of the matrix, in terms of the given constraint invokes information theory and, in particular, the procedure of maximising the entropy. The branching ratio is readily computed from the elements of the probability matrix. Very good agreement is obtained between the trajectory-computed and the synthetic branching ratio for all initial rotational states of HD.The F+HD reaction has three possible final arrangement channels (one nonreactive and two reactive ones) and is used to illustrate the structure of the reaction probability matrix and the associated entropy measures.     

The structure of plasma-polymerized toluene: A solid-state 13C-NMR study of isotopically labeled polymers

A detailed magic angle spinning ¹³C-NMR investigation of the intractable polymer prepared by plasma polymerization of toluene and isotopically labeled toluene led to a proposed model for the structure of the polymer and suggested some of the likely processes that occur in the gas phase leading to film formation. From the ¹³C spectra four resolved resonances permitted the determination of the contribution of nonprotonated and protonated unsaturated as well as methyl and other aliphatic carbons to the polymer structure. Specific ¹³C isotopic labeling of the methyl and phenyl C-1 toluene carbons in the injected liquid vapor allowed the destination of these carbons in the deposited polymer to be traced. The dominant structure is derived primarily from two precursors: benzyl radical and toluene itself. The ¹³C data further requires a net saturation of ca. 30% of the toluene double bonds and a net displacement of hydrogen by carbon on ca. 20% of the toluene ring carbons.     

Chemical shielding anisotropy of 13C and 15N in acetonitrile

High resolution ¹³C and ¹⁵N NMR spectra have been obtained for powdered CH₃CN. The presence of resolved dipolar structure in the ¹³C spectra permits the conclusion that the symmetry axis of the ¹³C shielding tensor lies along the CN bond direction.     

Enzymatic hydrogen atom abstraction from polyunsaturated fatty acids

The oxidation of polyunsaturated fatty acids such as arachidonic and linoleic acid initiates a plethora of cell signaling pathways in animals and plants. The chemistry of the enzymatic oxidation has been investigated for several enzymes, most notably prostaglandin synthase and the lipoxygenases, revealing many surprises and impressive examples of enzymatic control of hydrogen atom abstraction and subsequent oxygenation.     

A novel bioassay for screening and quantification of taxanes

We report a novel method for screening and quantifying tubulin-binding antimitotic agents that promote microtubule formation. The method is based on the shift in the peak of the fluorescence emission difference spectrum of tubulin complexed to metal free tetrakis(4-carboxyphenyl)porphyrin (TCPP) in the presence of antimitotic agents. Taxol (paclitaxel), an anti-tumor drug approved for the treatment of a variety of cancers, caused the appearance of a new fluorescence peak at 645 nm at concentrations as low as 125 nM, the intensity of which was a function of the paclitaxel concentration. Cephalomannine caused the induction of a new fluorescence peak at 651 nm only above 1 muM. Baccatin did not induce the appearance of any new peak within detectable operating measurement conditions. These observations are in accordance with the biological activities/cytotoxicities of these compounds. Accordingly, it is proposed that the new method can be used for high throughput screening of antimitotic compounds.     

Synthesis of persulfate containing poly (N-vinyl-2-pyrrolidone) (PVP) hydrogels in aqueous solutions by γ-induced radiation

The effect of ⁶⁰Co γ-irradiation on aqueous solutions of poly(N-vinyl-2-pyrrolidone) (PVP) in the presence of persulfate anion has been investigated. The gelation dose of PVP and persulfate containing PVP aqueous solutions has been determined. At low concentrations of persulfate (1.00–3.50%), gelation percentages exhibited a decreasing trend by increasing persulfate content in aqueous solutions of the polymer. The gelation doses of persulfate containing polymer solutions were calculated by the Charlesby–Pinner equation. It was observed that the gelation dose values were shifted to higher values by increasing persulfate concentration in solution. The ratio of the chain scission and crosslinking yields (G(s)/G(x)) was also determined. The results showed that the G(s)/G(x) ratios were smaller than one for PVP aqueous solution system, whereas those obtained for persulfate containing PVP aqueous solutions were higher than unity. The results implied that the chain scission of polymer is more effective than crosslinking in the presence of persulfate. Mechanism of the crosslinking and/or degradation and structure–property relationship of PVP and PVP/persulfate hydrogel systems were investigated by Fourier transformation infeared and thermal analysis (differential scanning calorimetry, thermal gravimetric analysis and differential thermai analysis) methods.     

Use of molecular dynamics in the design and structure determination of a photoinducible beta-hairpin

The study presented here consists of three parts. In the first, the ability of a set of differently substituted diazobenzene-based linkers to act as photoswitchable beta-turn building blocks was assessed. A 12-residue peptide known to form beta-hairpins was taken as the basis for the modeling process. The central (beta-turn) residue pair was successively replaced by six symmetrically ((o,o), (m,m), or (p,p)) substituted (aminomethyl/carboxymethyl or aminoethyl/carboxyethyl) diazobenzene derivatives leading to a set of peptides with a photoswitchable backbone conformation. The folding behavior of each peptide was then investigated by performing molecular dynamics simulations in water (4 ns) and in methanol (10 ns) at room temperature. The simulations suggest that (o,o)- and (m,m)-substituted linkers with a single methylene spacer are significantly better suited to act as photoswitchable beta-turn building blocks than the other linkers examined in this study. The peptide containing the (m,m)-substituted linker was synthesized and characterized by NMR in its cis configuration. In the second part of this study, the structure of this peptide was refined using explicit-solvent simulations and NOE distance restraints, employing a variety of refinement protocols (instantaneous and time-averaged restraining as well as unrestrained simulations). We show that for this type of systems, even short simulations provide a significant improvement in our understanding of their structure if physically meaningful force fields are employed. In the third part, unrestrained explicit-solvent simulations starting from either the NMR model structure (75 ns) or a fully extended structure (25 ns) are shown to converge to a stable beta-hairpin. The resulting ensemble is in good agreement with experimental data, indicating successful structure prediction of the investigated hairpin by classical explicit-solvent molecular dynamics simulations.