Insight into the Effect of Submicellar Concentrations of Sodium Deoxycholate on the Structure, Stability, and Activity of Bovine and Human Serum Albumin: An Interesting Comparison between Single and Double Tryptophan Proteins.

The progressive escalation in the applications of bile salts in diverse fields has triggered research on their interaction with various biological macromolecules, especially with proteins. A proper understanding of the interaction process of bile salts, particularly in the lower concentrations range, with the serum albumin seems important since the normal serum concentration of bile salts is approximately in the micromolar range. The current study deals with a comprehensive and comparative analysis of the interaction of submicellar concentrations of sodium deoxycholate (NaDC) with two homologous transport proteins: bovine serum albumin (BSA) and human serum albumin (HSA). HSA and BSA with one and two tryptophans, respectively, provide the opportunity for an interesting comparison of tryptophan fluorescence behavior on interaction with NaDC. The study suggests a sequential interaction of NaDC in three discrete stages with the two proteins. A detailed study using warfarin and ibuprofen as site markers provides information about the sites of interaction, which is further confirmed by inclusive molecular dynamics simulation analysis. Moreover, the comparison of the thermodynamics and stability of the NaDC-serum albumin complexes confirms the stronger interaction of NaDC with BSA as compared to that with HSA. The differential interaction between the bile salt and the two serum albumins is further established from the difference in the extent of decrease in the esterase-like activity assay of the proteins in the presence of NaDC. Therefore, the present study provides important insight into the effect of submicellar concentrations of NaDC on the structure, stability, and activity of the two homologous serum albumins and thus can contribute not only to the general understanding of the complex nature of serum albumin-bile salt interactions but also to the design of more effective pharmaceutical formulations in the field of drug delivery and biomedical research.

Photophysical Properties of Coumarin 1 in Bile Salt Aggregates: An Insight into the Role of Bile Salt Structure on the Aggregation Behavior.

The photophysical behavior of Coumarin 1 (Cou1), a well-known 7-aminocoumarin derivative, is very sensitive to the microenvironment in which it resides. In the present study, the effect of six bile salt variants on the photophysical behavior of Cou1 has been investigated. Dihydroxy (deoxycholates) as well as trihydroxy (cholates) bile salts with conjugated and unconjugated side chains have been chosen to get insight into the role of bile salt structure on the microenvironment of Cou1. Cou1 photophysics was found to be extremely sensitive to the aggregation process of the bile salt variants. The reduced polarity of the micellar environment stabilizes the planar intramolecular charge transferred state of Cou1, resulting in significant modulation in its photophysics in the bile salt media. The changes in the fluorescence parameters such as fluorescence intensity, emission energy, fluorescence quantum yield, anisotropy, and lifetime of Cou1 reveal that there is a distinct difference in the aggregation behavior of deoxycholates from that of cholates. The deoxycholates form micelles more or less critically similar to those of conventional surfactants, whereas the cholates self-assemble rather noncritically over a wide concentration range, thus signifying the vital role of the extra hydroxyl group in the aggregation pattern of trihydroxy bile salts. The conjugated bile salts are found to provide a relatively more compact, rigid, and hydrophobic microenvironment to Cou1 as compared to their unconjugated counterparts. Considering the significant modulation in the photophysical properties of Cou1, it has been employed as a molecular reporter for monitoring the aggregation process of bile salt variants and important information could be obtained about the effect of bile salt structure on the aggregation pattern and also about the micellar properties.

Spectroscopic investigation of interaction of Nile Blue A, a potent photosensitizer, with bile salts in aqueous medium.

Nile Blue A (NB) is one of the most studied benzophenoxazine dyes, as a potent photosensitizer for photodynamic therapy. The dye when administered intravenously disperses throughout the body by circulating through blood and is taken up by most cells that emphasize its interaction with various biomolecules. Therefore a rational understanding of the interaction of NB with relevant biological and biomimicking systems appears important. The focus of the present work is to investigate the interaction of NB with two bile salts sodium deoxycholate (NaDC) and sodium cholate (NaC) by spectroscopic techniques. The bile salts, in their premicellar concentration range, induce NB dimerization. Both H- and J-dimers are formed, however a major contribution is from the H-dimers. The extent of NB dimerization in NaDC, the dihydroxy bile salt, is higher than that in NaC, the trihydroxy bile salt. The bile salts when present above their micellar concentrations solubilize NB in its monomeric form. NB exhibits stronger binding and partitioning efficiency toward NaDC than NaC micelles at a given temperature. Binding and partitioning of NB to these micelles are spontaneous and exothermic in nature and these are enthalpy-driven processes. The spectral profiles and thermodynamic parameters of NB point toward the dissimilar nature of its environment in the micelles formed by the above two bile salts.