Spectroscopic, Zeta-potential and Surface Plasmon Resonance analysis of interaction between potential anti-HIV tannins with different flexibility and human serum albumin.

Tannins belong to secondary metabolites of plants that exhibit a variety of biological activities, including antiviral one. In this research, we studied the interaction of human serum albumin (HSA) with two ellagitannins: 2,4-valoneoyl-3,6-hexahydroxydiphenoyl-ß-d-glucose (T1) and 1,2-di-O-galloyl-3,6-valoneoyl-ß-d-glucose (T2) from Euphorbia species having antiviral potential against HIV and differing in molecular flexibility due to the presence of valoneoyl- and hexahydroxydiphenoyl groups. A fluorescence analysis demonstrated that the tannins studied strongly interacted with HSA and quenched tryptophan (Trp) fluorescence in the range of 0.25-4 µM. The quenching occurred by a static mechanism. The logKb for more flexible T2 was generally higher in comparison with stiffer T1 (4.94 ±â€¯0.82 vs. 4.12 ±â€¯0.31 and 4.94 ±â€¯0.53 vs. 4.07 ±â€¯0.45 for 296 K and 303 K respectively). The difference was also in the nature of the forces participating in the interaction with HSA. The stiff T1 reacted with HSA via hydrophobic forces, whereas the flexible T2 interacted with the protein by van der Waals forces and hydrogen bonds. The nature of the bonds was also confirmed by a study of the hydrophobicity of the compounds. Zeta-potential measurements showed slightly modifications of albumin electric charge but without significant changes in the surface structure of protein. Surface Plasmon Resonance imaging (SPRi) revealed that the used tannins fully saturated a 3 ng/mL solution of albumin at the concentrations of above 15 ng/mL. Our experiments clearly showed that the tannins used formed complexes with HSA and that the flexibility of the tannins was an important factor determining their interaction with the protein.

Comparative analysis of BPA and HQ toxic impacts on human erythrocytes, protective effect mechanism of tannins (Rhus typhina).

Several studies reported that bisphenol A (BPA) and its metabolite hydroquinone (HQ) have adverse effects on human and animal health. In this work, a comparative study of influence of the BPA and HQ, environment pollutants, on human erythrocytes was carried out. It was shown that BPA and HQ to varying extents caused oxidative damage in human erythrocytes: hemolysis, decreased GSH level, and methemoglobin formation. It was demonstrated that hydrolysable tannins 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (C55H40O34) and 1,2,3,4,6-penta-O-galloyl-ß-D-glucose (C41H32O26) (PGG) isolated from the Rhus typhina L. leaves in the range of 1-50 µM concentrations inhibited hemolysis and methemoglobin formation and also increased intracellular reduced glutathione in erythrocytes treated with BPA or HQ. It was revealed by electron paramagnetic resonance (EPR) using 5-doxyl-stearic acid (5-DS) that C55H40O34 and C41H32O26 increased the rigidity of erythrocyte membranes at the depth of 5th carbon atom of the fatty acid hydrocarbon chain. Taken together, these results allow to conclude that tannins from the Rhus typhina L. leaves protect erythrocytes from oxidative stress caused by BPA or HQ both due to their antioxidant activity as well as their interaction with the erythrocyte membrane components.

Interaction of α-synuclein with Rhus typhina tannin - Implication for Parkinson's disease.

The etiology of Parkinson's disease (PD) relates to α-synuclein, a small protein with the ability to aggregate and form Lewy bodies. One of its prevention strategies is inhibition of α-synuclein oligomerization. We have investigated the interaction of α-synuclein and human serum albumin with 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose (a tannin isolated from the plant Rhus typhina). Using fluorescence spectroscopy method we found that this tannin interacts strongly with α-synuclein forming complexes. Circular dichroism analysis showed a time-dependent inhibition of α-synuclein aggregation in the presence of the tannin. On the other hand, 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose had a much stronger interaction with human serum albumin than α-synuclein. The calculated binding constant for tannin-protein interaction was considerably higher for albumin than α-synuclein. This tannin interacted with albumin through a "sphere of action" mechanism. The results lead to the conclusion that 3,6-bis-О-di-О-galloyl-1,2,4-tri-О-galloyl-ß-d-glucose is a potent preventive compound against Parkinson's disease. However, this tannin interacts very strongly with human serum albumin, significantly reducing the bioavailability of this compound.

Specificity of Hydrolysable Tannins from Rhus typhina L. to Oxidants in Cell and Cell-Free Models.

Polyphenols of plant origin with wide range of antiradical activity can prevent diseases caused by oxidative and inflammatory processes. In this study, we show using ESR method that the purified water-soluble extract from leaves of Rhus typhina L. containing hydrolysable tannins and its main component, 3,6-bis-O-di-O-galloyl-1,2,4-tri-O-galloyl-ß-D-glucose (C55H40O34), displayed a strong antiradical activity against the synthetic 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) in homogenous (solution) and heterogeneous systems (suspension of DPPH containing liposomes) in the range of 1-10 µg/ml. The C55H40O34 and extract at 1-30 µg/ml also efficiently, but to a various degree, decreased reactive oxygen and nitrogen species (RONS) formation induced in erythrocytes by oxidants, following the sequence: tert-butyl hydroperoxide (tBuOOH) > peroxynitrite (ONOO-) >hypochlorous acid (HClO). The explanation of these differences should be seen in the specificity of scavenging different RONS types. These relationships can be represented for C55H40O34 and the extract by the following order of selectivity: O.-2 ≥ NO· > ·OH > 1O2. The extract exerted a more pronounced antiradical effect in reaction with DPPH and ROS in all models of oxidative stress in erythrocytes in comparison with C55H40O34. The redox processes between the extract components and their specificity in relation to RONS can underlie this effect.

Biomolecular Interactions of Tannin Isolated from Oenothera gigas with Liposomes.

We have examined the interaction between hydrolysable tannin 1-O-galloyl-4,6-hexahydroxydiphenoyl-ß-D-glucose (OGßDG) with neutral liposomes as a model of cell membranes composed of three lipids: lecithin, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) at different mass ratios. OGßDG in the concentration range 0.5-15 µg/ml (0.4-12 µM) strongly interacts with liposomal membranes by changing their structure, surface charge and fluidity. Used OGßDG molecules decrease and increase the rigidity of hydrophilic surface and hydrophobic parts of liposomes, respectively. At higher concentrations of tannin (>15 µM), liposomes are aggregated. Fourier Transform Infra-Red (FTIR) analysis showed that mainly -OH groups from OGßDG and also PO(2-) groups from phospholipids are responsible for the interaction. Obtained data indicate the importance of membrane lipid composition in interactions between tannins and cells.

Biophysical studies of interaction between hydrolysable tannins isolated from Oenothera gigas and Geranium sanguineum with human serum albumin.

Tannins, secondary plant metabolites, possess diverse biological activities and can interact with biopolymers such as lipids or proteins. Interactions between tannins and proteins depend on the structures of both and can result in changes in protein structure and activity. Because human serum albumin is the most abundant protein in plasma and responsible for interactions with important biological compounds (e.g. bilirubin) and proper blood pressure, therefore, it is very important to investigate reactions between HSA and tannins. This paper describes the interaction between human serum albumin (HSA) and two tannins: bihexahydroxydiphenoyl-trigalloylglucose (BDTG) and 1-O-galloyl-4,6-hexahydroxydiphenoyl-ß-d-glucose (OGßDG), isolated from Geranium sanguineum and Oenothera gigas leafs, respectively. Optical (spectrofluorimetric) and chiral optical (circular dichroism) methods were used in this study. Fluorescence analysis demonstrated that OGßDG quenched HSA fluorescence more strongly than BDTG. Both OGßDG and BDTG formed complexes with albumin and caused a red shift of the fluorescence spectra but did not significantly change the protein secondary structure. Our studies clearly demonstrate that the tested tannins interact very strongly with human serum albumin (quenching constant K=88,277.26±407.04 M(-1) and K=55,552.67±583.07 M(-1) respectively for OGßDG and BDTG) in a manner depending on their chemical structure.

Role of structural changes induced in biological membranes by hydrolysable tannins from sumac leaves (Rhus typhina L.) in their antihemolytic and antibacterial effects.

In this study, we found that the sumac tannins (Rhus typhina L.) exert to a various extent antihemolytic effects and antibacterial activity against Bacillus cereus and Pseudomonas aeruginosa depending on structural specificity of bacteria and different mechanisms of their toxic action. The sumac tannins exert the most expressed activity against B. cereus. The antihemolytic effect of the sumac tannins seems to be connected to a greater extent with their modifying action on the erythrocyte membrane structure. It was found that the sumac tannins are incorporated into the erythrocyte membrane, causing transformation of discocytes into echinocytes and enhancing the rigidity of the hydrophilic region of the lipid bilayer. We suggest that the embedding of sumac tannins into the membrane of erythrocytes alters their physical properties and, as a consequence, can limit their interaction with bacterial toxins.

Stabilization of erythrocytes against oxidative and hypotonic stress by tannins isolated from sumac leaves (Rhus typhina L.) and grape seeds (Vitis vinifera L.).

Erythrocytes are constantly exposed to ROS due to their function in the organism. High tension of oxygen, presence of hemoglobin iron and high concentration of polyunsaturated fatty acids in membrane make erythrocytes especially susceptible to oxidative stress. A comparison of the antioxidant activities of polyphenol-rich plant extracts containing hydrolysable tannins from sumac leaves (Rhus typhina L.) and condensed tannins from grape seeds (Vitis vinifera L.) showed that at the 5-50 µg/ml concentration range they reduced to the same extent hemolysis and glutathione, lipid and hemoglobin oxidation induced by erythrocyte treatment with 400 µM ONOO(-) or 1 mM HClO. However, extract (condensed tannins) from grape seeds in comparison with extract (hydrolysable tannins) from sumac leaves stabilized erythrocytes in hypotonic NaCl solutions weakly. Our data indicate that both hydrolysable and condensed tannins significantly decrease the fluidity of the surface of erythrocyte membranes but the effect of hydrolysable ones was more profound. In conclusion, our results indicate that extracts from sumac leaves (hydrolysable tannins) and grape seeds (condensed tannins) are very effective protectors against oxidative damage in erythrocytes.