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Reconstitution of energy transfer and electron transfer between solubilised pigment-protein complexes from thylakoid membranes. The role of acyl lipids
Authors:Denis J. Murphy
Affiliation:(1) Research Institute for Photosynthesis, University of Sheffield, 26 Taptonville Road, S10 5BR Sheffield, UK;(2) Department of Botany, University of Durham, South Road, DH1 3LE Durham, UK
Abstract:Solubilisation of thylakoid membranes from young leaves of Pisum sativum in the presence of Triton X-100 resulted in an almost complete loss of quenching of light-harvesting chlorophyll-protein (LHCP) fluorescence, as measured at 77°K. There were concomitant changes in the kinetics of light-saturation curves of electron transport from 2,6-dichlorophenolindophenol/ascorbate to methyl viologen. These effects were accompenied by a physical dissociation of LHCP polypeptides from photosystem I (PSI) and photosystem II (PSII) polypeptides, as determined by polyacrylamide gel-electrophoresis. Detergent-dialysis in the presence of exogenous purified galactolipids, about 80% of which were linoleoyl molecular species, only partially reversed these effects. However, detergent-dialysis using the phospholipids, phosphatidylglycerol and phosphatidylcholine, resulted in the substantial restoration of 77°K fluorescence quenching and the restoration of both emission spectra and electron transport kinetics of both Photosystems I and II that were typical of native membranes.Abbreviations Chl chlorophyll - DCPIP 2,6-dichlorophenolindophenol - DGD digalactosyldiacylglycerol - LHCP light-harvesting chlorophyll-protein - MGD monogalactosyldiacylglycerol - PCi phosphatidylcholine — Sigma grade NS - PCii beta-oleoyl, gamma-palmitoyl phosphalidylcholine - PG phosphatidylglycerol - PSI photosystem I - PSII photosystem II
Keywords:acyl lipid  chlorophyll-protein complexes  detergent-dialysis  energy transfer  reconstitution  thylakoid
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