Article

Kinetics of violaxanthin de-epoxidation by violaxanthin de-epoxidase, a xanthophyll cycle enzyme, is regulated by membrane fluidity in model lipid bilayers.

Department of Plant Physiology and Biochemistry, The Jan Zurzycki Institute of Molecular Biology and Biotechnology, Jagiellonian University, Kraków, Poland.
European Journal of Biochemistry (Impact Factor: 3.58). 10/2002; 269(18):4656-65. DOI: 10.1046/j.1432-1033.2002.03166.x
Source: PubMed

ABSTRACT This paper describes violaxanthin de-epoxidation in model lipid bilayers. Unilamellar egg yolk phosphatidylcholine (PtdCho) vesicles supplemented with monogalactosyldiacylglycerol were found to be a suitable system for studying this reaction. Such a system resembles more the native thylakoid membrane and offers better possibilities for studying kinetics and factors controlling de-epoxidation of violaxanthin than a system composed only ofmonogalactosyldiacylglycerol and is commonly used in xanthophyll cycle studies. The activity of violaxanthin de-epoxidase (VDE) strongly depended on the ratio of monogalactosyldiacylglycerol to PtdCho in liposomes. The mathematical model of violaxanthin de-epoxidation was applied to calculate the probability of violaxanthin to zeaxanthin conversion at different phases of de-epoxidation reactions. Measurements of deepoxidation rate and EPR-spin label study at different temperatures revealed that dynamic properties of the membrane are important factors that might control conversion of violaxanthin to antheraxanthin. A model of the molecular mechanism of violaxanthin de-epoxidation where the reversed hexagonal structures (mainly created by monogalactosyldiacylglycerol) are assumed to be required for violaxanthin conversion to zeaxanthin is proposed. The presence of monogalactosyldiacylglycerol reversed hexagonal phase was detected in the PtdCho/monogalactosyldiacylglycerol liposomes membrane by 31P-NMR studies. The availability of violaxanthin for de-epoxidation is a diffusion-dependent process controlled by membrane fluidity. The significance of the presented results for understanding themechanism of violaxanthin de-epoxidation in native thylakoid membranes is discussed.

0 Bookmarks
 · 
70 Views
  • [Show abstract] [Hide abstract]
    ABSTRACT: In their natural environment plants and algae are exposed to rapidly changing light conditions and light intensities. Illumination with high light intensities has the potential to overexcite the photosynthetic pigments and the electron transport chain and thus induce the production of toxic reactive oxygen species (ROS). To prevent damage by the action of ROS, plants and algae have developed a multitude of photoprotection mechanisms. One of the most important protection mechanisms is the dissipation of excessive excitation energy as heat in the light-harvesting complexes of the photosystems. This process requires a structural change of the photosynthetic antenna complexes that are normally optimized with regard to efficient light-harvesting. Enhanced heat dissipation in the antenna systems is accompanied by a strong quenching of the chlorophyll a fluorescence and has thus been termed non-photochemical quenching of chlorophyll a fluorescence, NPQ. The general importance of NPQ for the photoprotection of plants and algae is documented by its wide distribution in the plant kingdom. In the present review we will summarize the present day knowledge about NPQ in higher plants and different algal groups with a special focus on the molecular mechanisms that lead to the structural rearrangements of the antenna complexes and enhanced heat dissipation. We will present the newest models for NPQ in higher plants and diatoms and will compare the features of NPQ in different algae with those of NPQ in higher plants. In addition, we will briefly address evolutionary aspects of NPQ, i.e. how the requirements of NPQ have changed during the transition of plants from the aquatic habitat to the land environment. We will conclude with a presentation of open questions regarding the mechanistic basis of NPQ and suggestions for future experiments that may serve to obtain this missing information.
    Journal of Plant Physiology. 01/2014;
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Physical properties of thylakoid membranes isolated from barley were investigated by the electron paramagnetic resonance (EPR) spin labeling technique. EPR spectra of stearic acid spin labels 5-SASL and 16-SASL were measured as a function of temperature in secondary barley leaves during natural and dark-induced senescence. Oxygen transport parameter was determined from the power saturation curves of the spin labels obtained in the presence and absence of molecular oxygen at 25 °C. Parameters of EPR spectra of both spin labels showed an increase in the thylakoid membrane fluidity during senescence, in the headgroup area of the membrane, as well as in its interior. The oxygen transport parameter also increased with age of barley, indicating easier diffusion of oxygen within the membrane and its higher fluidity. The data are consistent with age-related changes of the spin label parameters obtained directly by EPR spectroscopy. Similar outcome was also observed when senescence was induced in mature secondary barley leaves by dark incubation. Such leaves showed higher membrane fluidity in comparison with leaves of the same age, grown under light conditions. Changes in the membrane fluidity of barley secondary leaves were compared with changes in the levels of carotenoids and proteins, which are known to modify membrane fluidity. Determination of total carotenoids and proteins showed linear decrease in their level with senescence. The results indicate that thylakoid membrane fluidity of barley leaves increases with senescence; the changes are accompanied with a decrease in the content of carotenoids and proteins, which could be a contributing factor.
    Journal of Plant Physiology. 01/2014;
  • [Show abstract] [Hide abstract]
    ABSTRACT: The physiological status of Cystoseira compressa, Padina pavonica and Palisada tenerrima was studied by in vivo chlorophyll fluorescence, pigment content, stoichiometry (C:N), accumulation of UV photoprotectors and antioxidant activity; comparing their photosynthetic response in a coastal lagoon (Mar Menor) and in Mediterranean coastal waters. In general, the specimens reached their highest ETRmax in spring in the Lagoon, but in summer in the Mediterranean, coinciding with their maximum biomass peak. The species exhibited a dynamic photoinhibition. Except C. compressa, they showed a lower decrease in Fv/Fm and higher recovery rates in the Mediterranean populations when exposed to high irradiance. The higher salinity and temperature of the lagoon could impair the photoprotection mechanisms. The acclimation to lagoon environments is species-specific and involves complex regulatory mechanisms. The results underline the importance of N in repair, avoidance, quenching and scavenging mechanisms. In general, Lagoon specimens showed higher pigment concentration. Although xanthophylls play important photo-protective and antioxidant roles, the observed trend is more likely to be explained by the higher temperatures reached in the lagoon compared to Mediterranean. Therefore the studied photosynthetic and biochemical mechanisms can be effective not only for high irradiance, but also for higher temperatures in a climate change scenario, but are highly dependent on nutrient availability.
    Marine environmental research 10/2014; · 2.34 Impact Factor

Full-text

Download
0 Downloads