Nucleic acid-metal interactions. IV. Complexes of Ag(I) with thymine and cytosine from studies of UV and IR dichroic spectra

The linear dichroism of the complexes of thymine and cytosine with silver(I) ions dispersed in stretched polyvinylalcohol films was measured in the infrared and in the near ultraviolet regions of the spectrum. The infrared results were helpful in establishing the orientation of the complexes in the films; they also confirmed that the site of silver binding in the case of cytosine involves the carbonyl oxygen. The ultraviolet spectra were deconvoluted into separate electronic transitions and the corresponding transition moments of silver complexes were determined.     

On the structure of the ethylene chlorine complex

An ethylene chlorine complex has been prepared in a nitrogen matrix at 20 K. Its ultraviolet and infrared spectra have been recorded. The results are compared with CNDO/2 calculations of structure. Both infrared data and CNDO calculations favor a C_2v symmetric complex. The CNDO calculations predict that the ClCl bond is orthogonal to the ethylene plane. The infrared data give no evidence for or against this prediction. Photolysis of the complex leads to the formation of 1,2-dichloroethane.     

Enzyme thermistors for process monitoring and control

In today’s biotechnology there is an increasing demand for appropriate analytical systems for process control. At present the most widely used control systems are based on measurements of pH, pO₂, and pCO₂. Such systems do not allow the direct measurement of substrates and products. To overcome this drawback sensors such as enzyme thermistors and enzyme electrodes have been designed and their development into industrial useful sensors for monitoring and controlling is the subject of active research.     

Microheterogeneity of human plasma lecithin: cholesterol acyltransferase examined by isoelectric focusing in immobilized pH gradients

The microheterogeneity of highly purified human plasma lecithin:cholesterol acyltransferase (LCAT) has been examined by electrophoresis in immobilized pH gradients in Immobiline-polyacrylamide gels of the pH ranges 4-7 and 4.2-4.9. Seven isoforms were obtained with LCAT isolated from pools of normal plasma. Using this technique the apparent pI values at 15 degrees C for the isoforms in the pH 4.2-4.9 gradient were 4.37, 4.42, 4.48, 4.53, 4.60, 4.67 and 4.74. (SD = +/- 0.03 for all). The most intensely stained band in the isoform pattern corresponded to the isoform with a pI value of 4.48.     

Radical-mediated carboxylation of alkyl iodides with [11C]carbon monoxide in solvent mixtures

[reaction: see text] [carboxyl-(11)C]Carboxylic acids were prepared from alkyl iodides via photoinitiated radical reactions using 10(-)(8) mol of [(11)C]carbon monoxide in binary and ternary homogeneous solvent mixtures. Short- (isobutyric), medium-, and long-chain saturated fatty acids (heptadecanoic) were labeled with isolated decay-corrected radiochemical yields ranging from 55% to 70% in 5-7-min reactions. The conversion of [(11)C]carbon monoxide to products reached 80-90%. To obtain good yields in the reactions performed in water-acetonitrile and water-THF mixtures, the addition of tetrabutylammonium hydroxide or potassium hydroxide was essential. The carboxylation was efficient for primary and secondary alkyl iodides. The carboxylation of tertiary iodides was feasible for 1-iodoadamantane but not for tert-butyl iodide. The dependence of the radiochemical yields on reaction time, photoirradiation conditions, and organic and inorganic additives was studied. The method provides a one-step route to [carboxyl-(11)C]carboxylic acids; traditional methods, in contrast, would require several steps. For example, using the devised reaction conditions, 3.19 GBq of purified [1-(11)C]1,10-decanedicarboxylic acid (specific radioactivity 188 GBq/mumol) was obtained within 35 min of the end of 10 muAh bombardment. (1-(13)C)4-Phenylbutyric acid was synthesized using ((13)C)carbon monoxide for identifying the labeling position with (1)H and (13)C NMR.     

Enantioselective luminescence quenching of DNA light-switch [Ru(phen)2dppz]2+ by electron transfer to structural homologue [Ru(phendione)2dppz]2+

The quenching of the luminescence of [Ru(phen)(2)dppz](2+) by structural homologue [Ru(phendione)(2)dppz](2+), when both complexes are bound to DNA, has been studied for all four combinations of Delta and Lambda enantiomers. Flow linear dichroism spectroscopy (LD) indicates similar binding geometries for all the four compounds, with the dppz ligand fully intercalated between the DNA base pairs. A difference in the LD spectrum observed for the lowest-energy MLCT transition suggests that a transition, potentially related to the final localization of the excited electron to the dppz ligand in [Ru(phen)(2)dppz](2+), is overlaid by an orthogonally polarized transition in [Ru(phendione)(2)dppz](2+). This would be consistent with a low-lying LUMO of the phendione moiety of [Ru(phendione)(2)dppz](2+) that can accept the excited electron from [Ru(phen)(2)dppz](2+), thereby quenching the emission of the latter. The lifetime of excited Delta-[Ru(phen)(2)dppz](2+) is decreased moderately, from 664 to 427 ns, when bound simultaneously with the phendione complex to DNA. The 108 ns lifetime of opposite enantiomer, Lambda-[Ru(phen)(2)dppz](2+), is only shortened to 94 ns. These results are consistent with an average rate constant for electron transfer of approximately 1.10(6) s(-1) between the phenanthroline- and phendione-ruthenium complexes. At binding ratios close to saturation of DNA, the total emission of the two enantiomers is lowered equally much, but for the Lambda enantiomer, this is not paralleled by a decrease in luminescence lifetime. A binding isotherm simulation based on a generalized McGhee-von Hippel approach shows that the Delta enantiomer binds approximately 3 times stronger to DNA both for [Ru(phendione)(2)dppz](2+) and [Ru(phen)(2)dppz](2+). This explains the similar decrease in total emission, without the parallel decrease in lifetime for the Lambda enantiomer. The simulation also does not indicate any significant binding cooperativity, in contrast to the case when Delta-[Rh(phi)(2)bipy](3+) is used as quencher. The very slow electron transfer from [Ru(phen)(2)dppz](2+) to [Ru(phendione)(2)dppz](2+), compared to the case when [Rh(phi)(2)phen](3+) is the acceptor, can be explained by a much smaller driving free-energy difference.     

Monitoring the DNA binding kinetics of a binuclear ruthenium complex by energy transfer: evidence for slow shuffling

The semirigid binuclear ruthenium complex Delta,Delta-[mu-(11,11'-bidppz)(phen)(4)Ru(2)](4+) has been shown to rearrange slowly from an initial groove-bound nonluminescent state to a final intercalated emissive state by threading one of its bulky Ru(phen)(2) moieties through the DNA base stack. When this complex binds to poly[d(A-T)(2)], a further increase in emission from the complex is observed after completion of the intercalation, assigned to reorganization of the intercalated complex. We here report a study of the threading process in poly[d(A-T)(2)], in which the minor groove binding dye DAPI is used as an energy transfer probe molecule to assess the distribution of ruthenium complex during and also after the actual threading phase. The emission from DAPI is found to change with the same rate as the emission from the ruthenium complex, and furthermore, DAPI does not disturb the binding kinetics of the latter, justifying it as a good probe of both the threading and the reorganization processes. We conclude from the change in the emission from both DAPI and the ruthenium complex with time that DAPI-ruthenium interactions are most pronounced during the process of threading of the complex, suggesting that the complexes are initially threaded slightly anticooperatively and thereafter redistribute along the DNA to reach their thermodynamically most favorable distribution. The final distribution is characterized by a small but significant binding cooperativity, probably as a result of hydrophobic interactions between the complex ions despite their tetravalent positive charges. The mechanism of "shuffling" the complex along the DNA chain is discussed, i.e., whether the ruthenium complex remains threaded (requiring sequential base-pair openings) or if unthreading followed by lateral diffusion within the ionic atmosphere of the DNA and rethreading occurs.     

Distribution analysis of ultra-high molecular mass poly(ethylene oxide) containing silica particles by size-exclusion chromatography with dual light-scattering and refractometric detection

Two different size-exclusion chromatography (SEC) systems, connected in-line either to a low-angle light scattering (LALS) or to a multiangle light scattering (MALS) detector, are employed for determination of molecular mass distributions (MMD) of poly(ethylene oxide) (PEO) samples having a weight average molecular mass up to eight millions. The detrimental effect of the presence of strongly scattering silica particles in the samples on the light scattering signal can be eliminated using a suitable sample dissolution procedure utilizing silica solubility in aqueous mobile phase. The selection of flow-rate and sample concentration have a large impact on the obtained results. Hydrodynamic retardation phenomena and nonlinearity effects are shown to introduce severe errors in the molecular mass distributions unless flow-rate and sample concentration are kept at sufficiently low levels. Self-compensating ability of the dual detection in flow-rate effects is shown to be the main advantage here. A good agreement between the results obtained using LALS and MALS detection is found provided that a carefully selected angular extrapolation procedure is used in the case of MALS data. Thus, using carefully selected experimental conditions, SEC with light-scattering (LS) and refractometric detection proved to be an efficient technique for MMD characterisation also of ultra-high molecular mass (UHM) PEO polymers.