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Physical properties of lipid bilayer membranes: relevance to membrane biological functions.

100%
Subczynski W. K., Wisniewska A.
Acta Biochimica Polonica
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2000
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vol. 47
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issue 3
613-625
EN
Over the last 25 years one of us (WKS) has been investigating physical properties of lipid bilayer membranes. In 1991 a group led by WKS was organized into the Laboratory of Structure and Dynamics of Biological Membranes, the effective member of which is AW. Using mainly the electron paramagnetic resonance (EPR) spin-labeling method, we obtained unexpected results, which are significant for the better understanding of the functioning of biological membranes. We have developed a new pulse EPR spin-labeling method for the detection of membrane domains and evaluation of lipid exchange rates. This review will be focused on our main results which can be summarized as follows: (1) Unsaturation of alkyl chains greatly reduces the ordering and rigidifying effects of cholesterol although the unsaturation alone gives only minor fluidizing effects, as observed by order and reorientational motion, and rather significant rigidifying effects, as observed by translational motion of probe molecules; (2) Fluid-phase model membranes and cell plasma membranes are not barriers to oxygen and nitric oxide transport; (3) Polar carotenoids can regulate membrane fluidity in a way similar to cholesterol; (4) Formation of effective hydrophobic barriers to the permeation of small polar molecules across membranes requires alkyl chain unsaturation and/or the presence of cholesterol; (5) Fluid-phase micro-immiscibility takes place in cis-unsaturated phosphatidylcholine-cholesterol membranes and induces the formation of cholesterol-rich domains; (6) In membranes containing high concentrations of transmembrane proteins a new lipid domain is formed, with lipids trapped within aggregates of proteins, in which the lipid dynamics is diminished to the level of gel-phase.
2
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Structural aspects of the antioxidant activity of lutein in a model of photoreceptor membranes

81%
Wisniewska-Becker A., Nawrocki G., Duda M., Subczynski W. K.
Acta Biochimica Polonica
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2012
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vol. 59
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issue 1
119-124
EN
It was shown that in membranes containing raft domains, the macular xanthophylls lutein and zeaxanthin are not distributed uniformly, but are excluded from saturated raft domains and about ten times more concentrated in unsaturated bulk lipids. The selective accumulation of lutein and zeaxanthin in direct proximity to unsaturated lipids, which are especially susceptible to lipid peroxidation, could be very important as far as their antioxidant activity is concerned. Therefore, the protective role of lutein against lipid peroxidation was investigated in membranes made of raft-forming mixtures and in models of photoreceptor outer segment membranes and compared with their antioxidant activity in homogeneous membranes composed of unsaturated lipids. Lipid peroxidation was induced by photosensitized reactions using rose Bengal and monitored by an MDA-TBA test, an iodometric assay, and oxygen consumption (using EPR spectroscopy and the mHCTPO spin label as an oxygen probe). The results show that lutein protects unsaturated lipids more effectively in membranes made of raft-forming mixtures than in homogeneous membranes. This suggests that the selective accumulation of macular xanthophylls in the most vulnerable regions of photoreceptor membranes may play an important role in enhancing their antioxidant properties and ability to prevent age-related macular diseases (such as age-related macular degeneration (AMD)).
3
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Carotenoid - membrane interactions in liposomes: effect of dipolar, monopolar, and nonpolar carotenoids

81%
Wisniewska A., Widomska J., Subczynski W. K.
Acta Biochimica Polonica
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2006
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vol. 53
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issue 3
475-484
EN
Electron paramagnetic resonance (EPR) spin-labeling methods were used to study the effects of carotenoids on the physical properties of saturated phosphatidylcholine (PC) membranes to evaluate the contribution of the terminal hydroxyl groups of xanthophyll molecules to the carotenoid-membrane interaction. Effects of the dipolar, terminally dihydroxylated carotenoid lutein on membrane phase transition, fluidity, order, and polarity were compared with those of monopolar (β-cryptoxanthin) and nonpolar (β-carotene) carotenoids. These effects were monitored at the membrane center as a function of the amount of the carotenoid added to the sample and as a function of temperature for fluid-phase membranes. PC membranes with different thickness (from 12 to 22 carbons in alkyl chains) were used. Carotenoids shifted to lower temperatures and broadened the main phase transition of PC membranes. They decreased the membrane fluidity and increased the order of alkyl chains. Carotenoids also increased the hydrophobicity of the membrane interior. These effects were the strongest for lutein, significantly weaker for β-cryptoxanthin, and negligible for β-carotene. They decreased with the increase of the membrane thickness. Presented results suggest that anchoring of carotenoid molecules at the opposite membrane surfaces by polar hydroxyl groups is significant in enhancing their effects on membrane properties. This manuscript also shows the ability of EPR spin-labeling methods to monitor different membrane properties that can be applied in biotechnological studies with the use of liposomes.
4
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Can macular xanthophylls replace cholesterol in formation of the liquid-ordered phase in lipid-bilayer membranes?

81%
Subczynski W. K., Wisniewska-Becker A., Widomska J.
Acta Biochimica Polonica
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2012
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vol. 59
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issue 1
109-114
EN
Lateral organization of membranes made from binary mixtures of dimyristoylphosphatidylcholine (DMPC) or dipalmitoylphosphatidylcholine (DPPC) and macular xanthophylls (lutein or zeaxanthin) was investigated using the saturation-recovery (SR) EPR spin-labeling discrimination by oxygen transport (DOT) method in which the bimolecular collision rate of molecular oxygen with the nitroxide spin label is measured. This work was undertaken to examine whether or not lutein and zeaxanthin, macular xanthophylls that parallel cholesterol in its function as a regulator of both membrane fluidity and hydrophobicity, can parallel other structural functions of cholesterol, including formation of the liquid-ordered phase in membranes. The DOT method permits discrimination of different membrane phases when the collision rates (oxygen transport parameter) differ in these phases. Additionally, membrane phases can be characterized by the oxygen transport parameter in situ without the need for separation, which provides information about the dynamics of each phase. In gel-phase membranes, two coexisting phases were discriminated in the presence of macular xanthophylls - namely, the liquid-ordered-like and solid-ordered-like phases. However, in fluid-phase membranes, xanthophylls only induce the solitary liquid-ordered-like phase, while at similar concentrations, cholesterol induces coexisting liquid-ordered and liquid-disordered phases. No significant differences between the effects of lutein and zeaxanthin were found.
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