In vitro reassociation of phycobiliproteins and membranes to form functional membrane-bound phycobilisomes |
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Authors: | Diana Kirilovsky Martin Kessel Itzhak Ohad |
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Institution: | 1. Department of Biological Chemistry, Institute of Life Sciences, The Hebrew University of Jerusalem, 91904 Jerusalem, Israel;2. Department of Membrane and Ultrastructure Research, Hebrew University - Hadassah Medical School, Jerusalem Israel |
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Abstract: | A membrane-bound phycobilisome complex has been isolated from the cyanobacterium Fremyella diplosiphon grown in green light, thus containing phycoerythrin in addition to phycocyanin and allophycocyanin. The complex was dissociated by lowering the salt concentration. In the mixture obtained, no energy transfer from phycoerythrin to chlorophyll (Chl) a was observed. Reassociation of the phycobiliproteins and membrane mixture was carried out by a gradual increase of the salt concentration. The complex obtained after reassociation was characterized by polypeptide composition, absorbance and fluorescence emission spectra and electron microscopy. These analyses revealed similar composition and structure for the original and reconstituted membrane-bound phycobilisomes. Fluorescence emission spectra and measurements of Photosystem II activity demonstrated energy transfer from phycoerythrin to Chl a (Photosystem II) in the reconstituted complex. Reassociation of mixtures with varying phycoerythrin / Chl ratio showed that the phycobiliprotein concentration was critical in the reassociation process. Measurements of the amount of phycobilisomes reassociated with the photosynthetic membrane did not show saturation of binding when increasing the phycobiliprotein concentration. The ratio phycoerythrin / Chl a in the native complex was 7:1 (mg / mg). When the phycobiliprotein concentration was increased during the reassociation process, a ratio of 13–15 mg phycoerythrin / mg Chl a could be obtained. Under these conditions, only part of the phycobilisomes attached to the thylakoids was able to transfer energy to Photosystem II. |
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Keywords: | Phycobiliprotein Phycobilisome Energy transfer Membrane reconstitution Photosystem II (Fremyella diplosiphon) Chl chlorophyll PS photosystem DCIP 2 6-dichlorophenolindophenol LDS lithium dodecyl sulfate |
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