Glass-forming tendency in the system water-dimethyl sulfoxide

The glass-forming tendency on cooling and the stability of the wholly amorphous state on warming have been previously reported for many cryoprotective solutions. However, unlike the other solutions, those of dimethyl sulfoxide (Me(2)SO) have not been studied on cooling. In this paper, the glass-forming tendency of Me(2)SO aqueous solutions has been measured for solutions containing 40, 43, 45, and 47.5% (w/w) Me(2)SO. At a concentration of 45% (w/w), the glass-forming tendency decreases in the following order: levo-2, 3-butanediol, 1,3-butanediol, 1,2-propanediol, 1,2,3-butanetriol, dimethyl sulfoxide, dimethylformamide, diethylformamide, 1, 4-butanediol, ethylene glycol, glycerol, 1,3-propanediol. New measurements have also been made on warming the Me(2)SO solutions.     

Dimethyl sulfoxide: A solvent for cytokinins in theAmaranthus bioassay

Dimethyl sulfoxide present in the agar medium at concentration 0.2 % (v/v) and lower does not inhibit cytokinin-induced betacyanin synthesis in theAmaranthus caudatus seedlings. The activity of kinetin, N⁶-(Δ²-isopentenyl)adenine andtrans- zeatin is the same when these cytokinins are dissolved in either water or dimethyl sulfoxide and incorporated into the medium after autoclaving. A simple method is described which allows the cytokinin activity of slightly water-soluble and thermolabile compounds,e.g. aromatic urea and thiourea derivatives, to be determined in theAmaranthus bioassay.     

Ion-molecule condensation reactions: A mechanism for organic synthesis in ionized reducing atmospheres

The CH₃ ⁺ ion, formed in ionized methane, undergoes consecutive eliminative condensation reactions with methane to form the carbonium ions C₂H₅ ⁺, i-C₃H₇ ⁺ and t-C₄H₉ ⁺. AtT<500°K, \(N_{CH_4 } \) ?10¹⁶ cm^?3 these ions react with NH₃ in competitive condensation-H⁺ transfer reactions, e.g. $$\begin{gathered} C_2 H_5 ^ + + NH_3 \xrightarrow{M} C_2 H_5 NH_3 ^ + \hfill \\ - - - \to NH_4 ^ + + C_2 H_4 \hfill \\ \end{gathered} $$ At particle densities of \(N_{CH_4 } \) <10¹⁶ cm^?3 proton transfer is the only significant reaction channel. At \(N_{CH_4 } \) >10¹⁷ cm^?3 condensation constitutes 5–20% of the overall reactions. The product of the condensation reaction further associates with CO₂ to form C₂H₅NH₃ ⁺·CO₂; the atomic composition of this cluster ion is identical with the protonated amino acid alanine. The carbonium ions i-C₃H₇ ⁺ and t-C₄H₉ ⁺ condense also with HCN to yield protonated isocyanides. HCNH⁺ also appears to condense with HCN atT>570°K, and form cluster ions with HCN at lower temperatures. The rate constants of the condensation reactions vary with temperature and pressure in a complex manner. Under conditions similar to those on Titan at an altitude of 100 km (T=100–150°K, \(N_{CH_4 } \) ≈10¹⁸ cm^?3), with a methane atmosphere containing 1% H₂ and traces of NH₃ and H₂O, ion-molecule condensation reactions followed by H⁺ transfer are expected to lead to the atmospheric synthesis of C₂H₆, C₃H₈, CH₃OH, C₂H₅OH and the terminal ions NH₄ ⁺, CH₃NH₃ ⁺ and C₂H₅NH₃ ⁺. At higher temperatures (250°K<T<400°K), the synthesis of i-C₄H₁₀, i-C₃H₇OH and t-C₄H₉OH and of the ions i-C₃H₇NH₃ ⁺ and t-C₄H₉NH₃ ⁺ is also expected. Electron recombination of the terminal ions may yield amines, imines and nitriles. Cycles of protonation and dissociative recombination of the alkanes and alcohols produced in condensation reactions will also produce unsaturated hydrocarbons, ketones and aldehydes in the ionized atmosphere.     

Resonance assignments of GTPase effector domain of dynamin in the aprotic solvent deuterated dimethyl sulfoxide

The GTPase effector domain (GED) is a subunit of dynamin, a multi-domain protein involved in endocytosis. GED forms a megadalton-sized self-assembly in vitro. The core of such huge assemblies is inaccessible to detailed Nuclear Magnetic Resonance characterization by conventional methods due to line broadening effects. Till date, there have been no studies to directly identify the residues involved in the core of the assembly. In this background we report here the NMR resonance assignments of deuterated dimethyl sulfoxide (DMSO-d6)-denatured GED from Homo sapiens. This will form the basis for probing the core of GED assembly and characterization of the association pathway driven by DMSO dilution.     

Methionine sulfoxide and the methionine sulfoxide reductase system as modulators of signal transduction pathways: a review

Methionine oxidation and reduction is a common phenomenon occurring in biological systems under both physiological and oxidative-stress conditions. The levels of methionine sulfoxide (MetO) are dependent on the redox status in the cell or organ, and they are usually elevated under oxidative-stress conditions, aging, inflammation, and oxidative-stress related diseases. MetO modification of proteins may alter their function or cause the accumulation of toxic proteins in the cell/organ. Accordingly, the regulation of the level of MetO is mediated through the ubiquitous and evolutionary conserved methionine sulfoxide reductase (Msr) system and its associated redox molecules. Recent published research has provided new evidence for the involvement of free MetO or protein-bound MetO of specific proteins in several signal transduction pathways that are important for cellular function. In the current review, we will focus on the role of MetO in specific signal transduction pathways of various organisms, with relation to their physiological contexts, and discuss the contribution of the Msr system to the regulation of the observed MetO effect.

     

Peroxidase-catalyzed N-demethylation reactions: deuterium solvent isotope effects

The effect of D2O on the kinetic parameters for the hydroperoxide-supported N-demethylation of N,N-dimethylaniline catalyzed by chloroperoxidase and horseradish peroxidase was investigated in order to assess the roles of exchangeable hydrogens in the demethylation reaction. The initial rate of the chloroperoxidase-catalyzed N-demethylation of N,N-dimethylaniline supported by ethyl hydroperoxide exhibited a pL optimum (where L denotes H or D) of 4.5 in both H2O and D2O. The solvent isotope effect on the initial rate of the chloroperoxidase-catalyzed demethylation reaction was independent of pL, suggesting that the solvent isotope effect is not due to a change in the pK of a rate-controlling ionization in D2O. The solvent isotope effect on the Vmax for the chloroperoxidase-catalyzed demethylation reaction was 3.66 +/- 0.62. In contrast, the solvent isotope effect on the Vmax for the horseradish peroxidase catalyzed demethylation reaction was approximately 1.5 with either ethyl hydroperoxide or hydrogen peroxide as the oxidant, indicating that the exchange of hydrogens in the enzyme and hydroperoxide for deuterium in D2O has little effect on the rate of the demethylation reaction. The solvent isotope effect on the Vmax/KM for ethyl hydroperoxide in the chloroperoxidase-catalyzed demethylation reaction was 8.82 +/- 1.57, indicating that the rate of chloroperoxidase compound I formation is substantially decreased in D2O. This isotope effect is suggested to arise from deuterium exchange of the hydroperoxide hydrogen and of active-site residues involved in compound I formation. A solvent isotope effect of 2.96 +/- 0.57 was observed on the Vmax/KM for N,N-dimethylaniline in the chloroperoxidase-catalyzed reaction.(ABSTRACT TRUNCATED AT 250 WORDS)     

Enzyme catalysed reactions in water - Organic solvent two-phase systems

Enzyme-catalysed transformations carried out in two-phase systems consisting of water and a water-immiscible organic solvent are discussed. The systems appear advantageous when substrates poorly soluble in water, such as steroids, are used. The methodology also makes possible the use of hydrolytic enzymes for the synthesis of ester bonds. Several applications of such two-phase systems are illustrated. The criteria that must be taken into consideration for selecting the most suitable organic solvents and the operational conditions are discussed.     

Chemical modification of silk sericin in lithium chloride/dimethyl sulfoxide solvent with 4-cyanophenyl isocyanate

This paper reports chemical modification of silk sericin in LiCl/dimethyl sulfoxide (DMSO) solvent with 4-cyanophenyl isocyanate. Sericin is a highly hydrophilic protein secreted by Bombyx mori, serving as a protein glue in a cocoon. LiCl/DMSO was found to be a good solvent of sericin and useful for homogeneous modification of its abundant hydroxyl groups under nonaqueous condition. Fourier transform infrared (FTIR) analysis of the modified sericins revealed that 4-cyanophenyl groups were incorporated into sericin molecules mainly through urethane linkages. Several characteristics of the modified sericins such as solubility characteristic, hygroscopic property, and thermal stability were investigated. Secondary structure analysis using FTIR spectra suggested that formation of strong intermolecular hydrogen bonds was inhibited by the modification that is probably attributable to the incorporation of bulky 4-cyanophenyl groups. These results demonstrate that chemical modification of sericin using LiCl/DMSO solvent markedly alters its characteristics.     

Chemiluminescent reactions in the Belousov-Zhabotinskii oscillating system

Chemiluminescence (CL) occurs during reactions of several of the components of the CH₂ (COOH)₂? KBrO₃? MnSO₄? H₂SO₄ self-sustained system. In contrast to the kinetics determined using potentiometry and spectrophotometry, the CL intensity kinetic curve during the oxidation of manganese (II) ions by acidic bromate has an extremum. Experimental dependences of the maximum CL intensity value on the initial reagent concentrations have been determined and compared with the results of the numerical simulation based on the commonly accepted Noyes-Field-Thompson mechanism.     

Baker''s yeast activity in an organic solvent system

The baker's yeast mediated reduction of ethyl 3-oxobutanoate-3-¹³C in hexane was conducted in an NMR tube at 20°C and a ¹³C NMR spectrum recorded each hour. A plot of relative peak intensity against time allowed the progress of the reaction to be monitored. A series of reactions was carried out in which the yeast was pretreated with the organic solvent system for 3, 6, 12 and 24 h prior to the addition of the substrate. From the initial rate of these reactions it was determined that in hexane the enzyme activity remained constant for about 12 h and then rapidly decreased until after 24 h very little activity remained. The reaction was also carried out at 10°C and 30°C. At the lower temperature, the reaction was slower but enzyme activity was maintained for more than 60 h, while at 30°C the enzyme activity had ceased after 8 h.     

Historical perspectives and the future of adverse reactions associated with haemopoietic stem cells cryopreserved with dimethyl sulfoxide

A retrospective review of the published literature identified several hundred adverse reactions (e.g. nausea, chills, cardiac arrhythmias, neurological symptoms and respiratory arrest) associated with the transplantation of stem cells cryopreserved with dimethyl sulfoxide. The occurrences of these are generally accepted as commonplace, as the majority of reactions are transient, whilst a few patients may require clinical treatment. This exploratory study is a collation of the historical data and the expectations for the notification of serious adverse reactions. Outline information is presented on the development of related European Directives, some technical aspects of dimethyl sulfoxide and the sequential stages of preservation and administration.     

Deuterium solvent isotope effect and proton-inventory studies of factor Xa-catalyzed reactions

Kinetic solvent isotope effects (KSIEs) for the factor Xa (FXa)-catalyzed activation of prothrombin in the presence and absence of factor Va (FVa) and 5.0 x 10(-5) M phospholipid vesicles are slightly inverse, 0.82-0.93, when substrate concentrations are at 0.2 Km. This is consistent with the rate-determining association of the enzyme-prothrombin assembly, rather than the rate-limiting chemical transformation. FVa is known to effect a major conformational change to expose the first scissile bond in prothrombin, which is the likely event triggering a major solvent rearrangement. At prothrombin concentrations > 5 Km, the KSIE is 1.6 +/- 0.3, when FXa is in a 1:1 ratio with FVa but becomes increasingly inverse, 0.30 +/- 0.05 and 0.19 +/- 0.04, when FXa/FVa is 1:4, with an increasing FXa and substrate concentration. The rate-determining step changes with the conditions, but the chemical step is not limiting under any circumstance. This corroborates the proposed predominance of the meizothrombin pathway when FXa is well-saturated with the prothrombin complex. In contrast, the FXa-catalyzed hydrolysis of N-alpha-Z-D-Arg-Gly-Arg-pNA.2HCl (S-2765) and H-D-Ile-L-Pro-L-Arg-pNA.HCl (S-2288) is most consistent with two-proton bridges forming at the transition state between Ser195 OgammaH and His57 N(epsilon)2 and His57 Ndelta1 and Asp102 COObeta- at the active site, with transition-state fractionation factors of phi1 = phi2 = 0.57 +/- 0.07 and phiS = 0.78 +/- 0.16 for solvent rearrangement for S-2765 and phi1 = phi2 = 0.674 +/- 0.001 for S-2288 under enzyme saturation with the substrate at pH 8.40 and 25.0 +/- 0.1 degrees C. The rate-determining step(s) in these reactions is most likely the cleavage of the C-N bond and departure of the leaving group.