Наукові роботи. Факультет радіофізики, біомедичної електроніки та комп’ютерних систем
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Документ Hemoglobin binding to phospholipid membranes as revealed by pyrene fluorescence study(Харьковский Национальный Университет им. В.Н.Каразина, 2011) Kutsenko, O.K.; Gorbenko, G.P.; Trusova, V.M.In this work hemoglobin (Hb) association with lipid bilayers was investigated using fluorescent probe pyrene. Model membranes were prepared from zwitterionic lipid phosphatidylcholine (PC), anionic lipid phosphatidylglycerol (PG) and cholesterol (Chol). Hb-lipid binding was followed by the pyrene fluorescence quenching. Hb-induced decrease of pyrene monomer fluorescence was followed by the increase of relative intensities of vibronic bands. Presumably, Hb penetration into the bilayer increases the space between neighbouring lipids and promotes water penetration into the membrane core. Pyrene excimer emission quenching was interpreted in terms of resonance energy transfer. The greatest depth of Hb penetration into the lipid bilayer was observed in PC vesicles. In Chol-containing liposomes sterol condensing effect prevents deep protein penetration into the membrane. PG has an ability to stabilize lipid bilayers due to the ordered state of its lipid tails and H-bonding interactions between lipid molecules. This also can prevent Hb access to the inner membrane regions.Документ Influence of oligomeric and fibrillar lysozyme on physical properties of model membranes(Харьковский Национальный Университет им. В.Н.Каразина, 2011) Kastorna, A.P.; Trusova, V.M.; Gorbenko, G.P.A pathological hallmark of more than 20 human diseases including Alzheimer’s disease, Parkinson’s disease, type II diabetes is the deposition in organs and tissues of insoluble highly ordered protein aggregates, called amyloid fibrils. It is becoming widely recognized that toxicity of amyloid species is related to their interactions with cell membranes. In the present study we focused our efforts on the examination of the influence of amyloid fibrils and their precursors (oligomeric aggregates) of lysozyme on the structural and physical properties of the model membranes composed of phosphatidylcholine and its mixture with cholesterol. For evaluating the extent of lipid bilayer modifications, we used fluorescence spectroscopy technique. The results of pyrene excimerization measurements showed that amyloid protein reduces membrane fluidity. Analysis of Laurdan emission spectra revealed the ability of lysozyme aggregates to produce bilayer dehydration. The most pronounced membrane-modifying effects were observed in the case of oligomeric lysozyme. Significantly less influence of pathogenic protein aggregates on the physical properties of cholesterol-containing vesicles confirmed the hypothesis on the preventive role of cholesterol in amyloid-related diseases.Документ Lipid bilayer modification induced by fibrillar lysozyme: fluorescence spectroscopy study(Харьковский Национальный Университет им. В.Н.Каразина, 2010) Kastorna, A.P.; Trusova, V.M.; Gorbenko, G.P.The correlation between neurodegenerative diseases (Parkinson’s, Alzheimer’s diseases), type II diabetes, systemic amyloidosis, etc. and deposition of protein aggregates in brain and other tissues has long been established. A growing body of evidence has demonstrated that binding of amyloid proteins to the membrane may underlie their cytotoxic effect. It was shown that amyloid toxicity arises primarily from a soluble oligomeric form (pre-fibrillar aggregates) of the peptide rather than amyloid monomers or mature fibrils. The molecular basis of the amyloid protein toxicity is not sufficiently clear and requires further investigation. In view of this, the present study has been undertaken to ascertain the effect of fibrillar aggregates of lysozyme on the structural and physical properties of model membranes (liposomes) composed of zwitterionic lipid phosphatidylcholine and its mixture with cholesterol (30 mol%). To this end, two fluorescent probes with different properties and bilayer location, pyrene and Laurdan, have been employed. Pyrene spectra have characteristic vibronic structure in the region of 370-400 nm. Relative intensities of vibronic transitions exhibit dependence on solvent polarity. Excited species of pyrene can interact with non-excited ones thus forming excited state dimers – excimers. Excimer-to-monomer fluorescence intensity ratio reflects the extent of pyrene excimerization, which depends mainly on the rate of monomer lateral diffusion in lipid bilayer, being a function of the density of lipid molecular packing. Analysis of pyrene emission spectra revealed the absence of any influence of fibrillar lysozyme on the structural state of bilayer acyl chain region. Laurdan is an amphiphilic fluorescent probe, whose emission spectra are sensitive to the environmental polarity (hydration level). In the solvents of high polarity, Laurdan shows a considerable shift of its emission spectrum to longer wavelengths due to the dipolar relaxation processes. The changes in the emission spectrum of Laurdan were characterized by the generalized polarization value (GP). In all types of liposomes increasing concentration of fibrillar lysozyme resulted in the increment of GP, suggesting that amyloid fibrils cause the decrease in the lipid bilayer polarity.