EPR characterization of an oxygen-evolving photosystem II preparation from the transformable cyanobacterium Synechocystis 6803.
The transformable cyanobacterium Synechocystis 6803 has a photosynthetic apparatus that is similar to that of plants. Because of the ease with which this organism can be genetically manipulated and isotopically labeled, Synechocystis has been used extensively in recent studies of electron transfer in the water-splitting complex, photosystem II. Here, we present the first EPR characterization of a highly active oxygen-evolving preparation from this organism. This preparation shows oxygen-evolution activities in the range from 2400-2600 mumol of O2/(mg of chlorophyll.h). We show that this preparation is stable enough for room temperature EPR studies. We then use this assay to show that the lineshapes of the D+ and Z+ tyrosine radicals are identical in this preparation, as has been observed in photosystem II complexes from a wide variety of photosynthetic species. We also present the first multiline EPR spectrum that has been observed from the Synechocystis manganese cluster.
Noren
G H
GH
Department of Biochemistry, University of Minnesota, St. Paul 55108.
Boerner
R J
RJ
Barry
B A
BA
D015153:Blotting, Western; D002734:Chlorophyll / Q000032:analysis; D000458:Cyanobacteria / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D004579:Electron Transport; D004591:Electrophoresis, Polyacrylamide Gel; D045342:Light-Harvesting Protein Complexes; D008345:Manganese / Q000032:analysis; D008970:Molecular Weight; D010100:Oxygen / Q000032:analysis; D010788:Photosynthesis; D045322:Photosynthetic Reaction Center Complex Proteins / Q000302:isolation & purification / Q000378:metabolism*; D045332:Photosystem II Protein Complex
D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; D045332:Photosystem II Protein Complex; D002734:Chlorophyll; D008345:Manganese; D010100:Oxygen