Characterization of an improved reaction center preparation from the photosynthetic green sulfur bacterium Chlorobium containing the FeS centers FA and FB and a bound cytochrome subunit.
A photosynthetic reaction center complex was prepared from the green sulfur bacterium Chlorobium by solubilization of chlorosome-depleted membranes with lauryl maltoside, followed by anion-exchange chromatography and molecular sieve chromatography. The purified complex was characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, optical spectroscopy, and EPR spectroscopy. The major bands migrated at apparent molecular masses of 50, 42, and 32 kDa (heme-staining) and additional weaker bands at 22, 15, and 12 kDa. The isolated reaction center complex contained about 40 bacteriochlorophyll alpha molecules per primary electron donor, P840, assayed by photooxidation. It was competent in stable low-temperature photoreduction of the FeS centers FA and FB. The spectra of these acceptors and their low-temperature photochemistry in the purified complex were the same as found in intact Chlorobium membranes and similar to what had been described for photosystem I from plants. Membrane-bound cytochrome c553 copurified with the reaction center complex. A ratio of about four hemes per P840 was determined. This result indicates that cytochrome c553 that is closely associated with the reaction center is a tetraheme cytochrome, as described for some purple bacteria.
Feiler
U
U
Molekulare Membranbiologie, Max-Planck-Institut für Biophysik, Frankfurt am Main, FRG.
Nitschke
W
W
Michel
H
H
D001429:Bacteriochlorophylls / Q000032:analysis; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003574:Cytochrome c Group / Q000302:isolation & purification* / Q000378:metabolism; D003902:Detergents; D004578:Electron Spin Resonance Spectroscopy; D004591:Electrophoresis, Polyacrylamide Gel; D005960:Glucosides; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D045342:Light-Harvesting Protein Complexes; D008970:Molecular Weight; D010777:Photochemistry; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012245:Rhodospirillales / Q000737:chemistry*; D013053:Spectrophotometry
D001429:Bacteriochlorophylls; D003574:Cytochrome c Group; D003902:Detergents; D005960:Glucosides; D007506:Iron-Sulfur Proteins; D045342:Light-Harvesting Protein Complexes; D045322:Photosynthetic Reaction Center Complex Proteins; C083951:cytochrome c553; C040358:dodecyl maltoside; C022302:cytochrome C-552