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The influence of phase transitions in phosphatidylethanolamine models on the activity of violaxanthin de-epoxidase
Authors:Astrid Vieler  Holger A Scheidt  Cindy Montag  Martin Lohr  Daniel Huster  Reimund Goss
Institution:a Institute of Biology I, University of Leipzig, Johannisallee 21-23, 04103 Leipzig, Germany
b Junior Research Group “Structural Biology of Membrane Proteins” Institute of Biochemistry/Biotechnology, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, D-06120 Halle, Germany
c Institute of General Botany, Johannes Gutenberg University of Mainz, Müllerweg 6, 55128 Mainz, Germany
d Institute of Medical Physics and Biophysics, University of Leipzig, D-04107 Leipzig, Germany
Abstract:In the present study, the influence of the phospholipid phase state on the activity of the xanthophyll cycle enzyme violaxanthin de-epoxidase (VDE) was analyzed using different phosphatidylethanolamine species as model lipids. By using 31P NMR spectroscopy, differential scanning calorimetry and temperature dependent enzyme assays, VDE activity could directly be related to the lipid structures the protein is associated with. Our results show that the gel (Lβ) to liquid-crystalline (Lα) phase transition in these single lipid component systems strongly enhances both the solubilization of the xanthophyll cycle pigment violaxanthin in the membrane and the activity of the VDE. This phase transition has a significantly stronger impact on VDE activity than the transition from the Lα to the inverted hexagonal (HII) phase. Especially at higher temperatures we found increased VDE reaction rates in the presence of the Lα phase compared to those in the presence of HII phase forming lipids. Our data furthermore imply that the HII phase is better suited to maintain high VDE activities at lower temperatures.
Keywords:Ax  Antheraxanthin  DOPE  dioleoylphosphatidylethanolamine  DSC  differential scanning calorimetry  DTT  dithiothreitol  EDTA  ethylenediaminetetraacetate  eggPE  phosphatidylethanolamine from egg yolk  GdmHCl  guanidinium hydrochloride  GSH  reduced glutathione  GSSG  oxidized glutathione  HII  inverted hexagonal phase  IPTG  isopropylthiogalactoside  Lα  liquid-crystalline phase  Lβ  gel phase  MGDG  monogalactosyldiacylglycerol  POPE  palmitoyloleoylphosphatidylethanolamine  VDE  violaxanthin de-epoxidase  Vx  Violaxanthin  Zx  Zeaxanthin
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