Studies on canthaxanthin in lipid membranes.

Polar carotenoid pigment - canthaxanthin - has been found to interfere with the organization of biological membranes, in particular of the retina membranes of an eye of primates. The organization of lipid membranes formed with dipalmitoylphosphatidylcholine (DPPC) and egg yolk phosphatidylcholine containing canthaxanthin was studied by means of several techniques including: electronic absorption spectroscopy, linear dichroism, X-ray diffractometry, (1)H-NMR spectroscopy and FTIR spectroscopy. It appears that canthaxanthin present in the lipid membranes at relatively low concentration (below 1 mol% with respect to lipid) modifies significantly physical properties of the membranes. In particular, canthaxanthin (i) exerts restrictions to the segmental molecular motion of lipid molecules both in the headgroup region and in the hydrophobic core of the bilayer, (ii) promotes extended conformation of alkyl lipid chains, (iii) modifies the surface of the lipid membranes (in particular in the gel state, L(beta )) and promotes the aggregation of lipid vesicles. It is concluded that canthaxanthin incorporated into lipid membranes is distributed among two pools: one spanning the lipid bilayer roughly perpendicularly to the surface of the membrane and one parallel to the membrane, localized in the headgroup region. The population of the horizontal fraction increases with the increase in the concentration of the pigment in the lipid phase. Such a conclusion is supported by the linear dichroism analysis of the oriented lipid multibilayers containing canthaxanthin: The mean angle between the dipole transition moment and the axis normal to the plane of the membrane was determined as 20+/-3 degrees at 0.5 mol% and 47+/-3 degrees at 2 mol% canthaxanthin. The analysis of the absorption spectra of canthaxanthin in the lipid phase and (1)H-NMR spectra of lipids point to the exceptionally low aggregation threshold of the pigment in the membrane environment (approximately 1 mol%). All results demonstrate a very strong modifying effect of canthaxanthin with respect to the dynamic and structural properties of lipid membranes.

Heat-induced and light-induced isomerization of the xanthophyll pigment zeaxanthin.

Zeaxanthin is a xanthophyll pigment that plays important physiological functions both in the plant and in the animal kingdom. All-trans is a stereochemical conformation of zeaxanthin reported as specific for the thylakoid membranes of the photosynthetic apparatus and the retina of an eye. On the other hand, the pigment is subjected, in natural environment, to the conditions that promote stereochemical isomerization, such as illumination and elevated temperature. In the present work, the light-induced and heat-induced (the temperature range 35-95 degrees C) isomerization of all-trans zeaxanthin in organic solvent environment has been analyzed by means of the HPLC technique. The 13-cis conformation has been identified as a major one among the isomerization products. The activation energy of the all-trans to 13-cis isomerization has been determined as 83 +/- 4 kJ/mol and the activation energy of the back reaction as 30 +/- 7 kJ/mol. The reaction of isomerization of the all-trans zeaxanthin at 25 degrees C was substantially more efficient upon illumination. Four different wavelengths of light have been selected for photo-isomerization experiments: 450, 540, 580 and 670 nm, corresponding to the electronic transitions of zeaxanthin from the ground state to the singlet excited states: 1(1)Bu+,3(1)Ag-,1(1)Bu- and 2(1)Ag-, respectively. The quantum efficiency of the all-trans zeaxanthin isomerization induced by light at different wavelengths: 450, 540, 580 and 670 nm was found to differ considerably and was in the ratio as 1:15:160:29. The sequence of the quantum efficiency values suggests that the carotenoid triplet state 1(3)Bu, populated via the internal conversion from the 1(3)Ag triplet state which is generated by the intersystem crossing from the 1(1)Bu- state may be involved in the light-induced isomerization. A physiological importance of the isomerization of zeaxanthin in the retina of an eye, photosynthetic apparatus and of the pigment active as a blue light photoreceptor in stomata is briefly discussed.

Interaction of isomeric forms of xanthophyll pigment zeaxanthin with dipalmitoylphosphatidylcholine studied in monomolecular layers.

Two-component monomolecular layers were formed with DPPC and two stereoisomers of zeaxanthin 9-cis and 13-cis at the argon-water interface. Very distinct over-additivity which represents affection of a lipid arrangement in the membrane has been observed in the case of zeaxanthin 9-cis (maximum at 20 mol%) but not in the case of zeaxanthin 13-cis. The differences in the organization of the isomers of zeaxanthin-DPPC monolayers are interpreted in terms of the different orientation of both xanthophylls at the interface observed at relatively high surface pressures (>25 mN/m) comparable to the surface pressures of biomembranes. The results are consistent with the model according to which zeaxanthin 9-cis adopts a vertical orientation at the polar-nonpolar interface in contrast to zeaxanthin 13-cis, which is oriented horizontally owing to the fact that it interacts by two hydroxyl groups with the same hydrophobic-hydrophilic interface in the monolayer. The findings are discussed in comparison with the behavior of zeaxanthin in the conformation all-trans in the same system. Zeaxanthin all-trans forms efficiently molecular aggregates in the mixed monolayers in contrast to cis isomers. Circular dichroism measurements show the formation of molecular structures by zeaxanthin 13-cis that are interpreted as dimers. FTIR measurements show that these dimers are stabilized by van der Waals interactions unlike aggregated structures formed by all-trans zeaxanthin that are stabilized by hydrogen bonding. Physiological importance of the differences in aggregation and orientation of stereoisomers of zeaxanthin in lipid environment is discussed.