Eva Valušová

Structural and thermodynamic behavior of cytochrome c assembled with glutathione-covered gold nanoparticles

The conformational changes of horse heart ferricytochrome c (cyt c) after association of gold nanoparticles have been studied by electronic absorption spectroscopy and circular dichroism (CD). Our results show that the structural stability around the heme of complexed cyt c was increased successfully. Glutathione-layered gold nanoparticles caused a significant increase of the apparent pK values of the cyt c alkaline transition. Similarly, the heme crevice became more stable to heat after assembly of cyt c with gold nanoparticles. In contrast, gold nanoparticles weaken the overall thermal stability of the cyt c by decreasing the denaturation temperature estimated from far-UV CD measurements. Similar behavior has previously been reported for cyt c complexed with physiological redox partners as well as hydrophilic polyanions.

Effect of N-domain on the stability of elongation factor Ts from Thermus thermophilus

Elongation factor Ts (EF-Ts) from Thermus thermophilus forms a stable, functionally active homodimer in solution. Its monomer is composed of two domains: amino-terminal domain containing 50 amino acid residues and a larger, 146 residues long, C-domain which participates in dimerization of EF-Ts. Effect of removal of the N-domain on the conformational stability of EF-Ts has been studied. For comparison, the stabilities of both the full-length EF-Ts and its C-domain were studied by differential scanning calorimetry, electronic absorption and fluorescence spectroscopies over a pH range from 4 to approximately 13. Thermal denaturation of EF-Ts and of C-domain, followed by circular dichroism at 222 nm, at pH 7.0, and the pH dependence of the fluorescence of the single tryptophan 30 residue indicate a conformational instability of the N-domain. While N-domain does not affect the stability of full-length EF-Ts at acidic pH, its removal leads to stabilization of the rest of the protein at basic pH. This is reflected by higher values of transition temperatures and calorimetric enthalpies of C-domain as compared to the full-length EF-Ts. High mobility of the N-domain in alkaline pH conditions decreased the thermal stability of covalently linked C-domain of EF-Ts. An increase in intramolecular interactions at acidic pH together with a decrease of conformational entropies of the thermally denatured proteins most likely diminishes this destabilization effect.

Water treatment using activated carbon supporting silver and magnetite.

Recent efforts in water purification have led to the development of novel materials whose unique properties can offer effective biocidal capabilities with greater ease of use and at lower cost. In this study, we introduce a novel procedure for the preparation of activated carbon (charcoal) composite in which magnetite and silver are incorporated (MCAG); we also describe the use of this material for the disinfection of surface water. The formation process of magnetic MCAG composite was studied using ultraviolet-visible spectroscopy. The results demonstrated the high sorption efficiency of AgNO₃ to magnetic activated carbon. The antimicrobial capabilities of the prepared MCAG were examined and the results clearly demonstrate their inhibitory effect on total river water bacteria and on Pseudomonas koreensis and Bacillus mycoides cultures isolated from river water. The bacterial counts in river water samples were reduced by five orders of magnitude following 30 min of treatment using 1 g l⁻¹ of MCAG at room temperature. The removal of all bacteria from the surface water samples implies that the MCAG material would be a suitable disinfectant for such waters. In combination with its magnetic character, MCAG would be an excellent candidate for the simple ambulatory disinfection of surface water.

Interactions between heparinoids and alcohol dehydrogenase

The interaction between polysulfated polysaecharides (low‐molecular‐weight heparin LMWH, dextran sulfate DS and pentosan sulfate PS) and yeast alcohol dehydrogenase (YADH) was investigated. The fluorescence and UV spectra of YADH after adding the tested polysaccharides have confirmed the interaction between the enzyme and these compounds. Kinetic studies have shown that LMWH, DS and PS are inhibitors of YADH (mixed type with respect to NAD). The most potent inhibitor is PS (ID50=37.5 ng/ml, Ki=0.6 μM). The inhibition effect depends on the ionic strength (the inhibition decreased by about 50% in the presence of 100 mM Na2SO4) and pH value (the inhibition decreased at pH>7). The results indicate that the inhibition effect of these polyanions is caused by their electrostatic interactions with the NAD‐binding region of YADH.

Excitation of triplet states of hypericin in water mediated by hydrotropic cromolyn sodium salt

Hypericin (Hyp) is a hydrophobic pigment found in plants of the genus Hypericum which exhibits low levels of solubility in water. This work shows that the solubility of Hyp can be significantly increased through the addition of cromolyn disodium salt (DSCG). Performed studies using UV-VIS absorption and fluorescence spectroscopies demonstrate that Hyp remains in a predominantly biologically photodynamic active monomeric form in the presence of DSCG at concentrations ranging from 4.6×10-3 to 1.2×10-1mol·L-1. The low association constant between Hyp and DSCG (Ka=71.7±2M-1), and the polarity value of 0.3 determined for Hyp in a DSCG-water solution, lead to a suggestion that the monomerization of Hyp in aqueous solution can be explained as a result of the hydrotropic effect of DSCG. This hydrotropic effect is most likely a result of interactions between two relative rigid aromatic rings of DSCG and a delocalized charge on the surface of the Hyp molecule. The triplet-triplet (T-T) electronic transition observed in is Hyp in the presence of DSCG suggests a possible production of reactive oxygen species once Hyp is irradiated with visible light in a DSCG aqueous solution.