The interactions of bovine serum albumin (BSA) with ionic surfactants (sodium dodecyl sulfate, SDS, and cetyltrimethylammonium bromide, CTAB) and β-cyclodextrin (β-CD) have been investigated by electron paramagnetic resonance (EPR) and circular dichroism measurements. The spin probe selected to report on the interaction of albumin with surfactants and/or β-CD was 4-N,N-dimethyl hexadecyl ammonium-2,2,6,6-tetramethylpiperidine-1-oxyl iodide (CAT16), on account of (a) its balance between electrostatic and hydrophobic character and (b) the ability of BSA to form complexes with various organic molecules. The distribution of the spin probe among different environments in solutions containing only BSA was confirmed by the existence of two components in the EPR spectra: one revealing a restricted mobility of the spin probe, attributed to the protein-spin probe complex, and another one showing free movement, attributed to the spin probe in solution. The presence of surfactants and/or β-CD alters the distribution of CAT16 between various compartments in each system. Formation of protein aggregates as a result of thermal denaturation was evidenced by the appearance of an immobilized component in the EPR spectrum. This component is not present in the EPR spectra of CAT16 in protein/surfactant or protein/cyclodextrin solutions. Circular dichroism spectra of BSA provided information about changes in the secondary structure of the protein induced by the presence of surfactants and/or cyclodextrin in solution. The results demonstrate that β-CD hinders the interaction between the employed surfactants and the protein. The cationic surfactant (CTAB) induces changes in protein conformation at a lower concentration compared to the anionic surfactant (SDS).

No abstract

No abstract

No abstract

Samples of human serum albumin (HSA) obtained as a result of heat denaturation followed by refolding controlled by a cooling of the protein solution were studied by several methods: chromatographic measurements, kinetic of the reaction with a water soluble free radical and by electron paramagnetic resonance (EPR) spectroscopy. In this context the interaction of this protein with β-cyclodextrin (β-CD) and sodium dodecyl sulfate (SDS) was also investigated. Reversed phase thin layer chromatography (RP-TLC) showed changes in lipophylicity of HSA, which are related with the existence of different ensembles of conformers. The UV-Vis absorption spectra had shown the broadening of absorption band of the protein and a hyperchrom effect in the presence of SDS; β-CD reduces the effect of SDS on protein UV-Vis spectra.Kinetic measurements related to the reaction of HSA with a water soluble DPPH type free radical provided evidence that reactivity of the HSA denaturated conformers is higher compared with the natural conformer. The affinity of SDS to the albumins surface and the effect of β-CD on the SDS/protein aggregates were also evident by changes in the EPR spectra of the spin probe CAT16.