ABSTRACT
A method is described which allows the selective release and removal of the Bchla-B800 molecules from the LH2 complex of Rhodopseudomonas acidophila 10050. This procedure also allows reconstitution of approximately 80% of the empty binding sites with native Bchla. As shown by circular dichroism spectroscopy, the overall structures of the B850-only and reconstituted complexes are not affected by the pigment-exchange procedure. The pigments reconstituted into the B800 sites can also efficiently transfer excitation energy to the Bchla-B850 molecules.
Subject(s)
Bacterial Proteins , Bacteriochlorophylls/chemistry , Bacteriochlorophylls/metabolism , Light-Harvesting Protein Complexes , Photosynthetic Reaction Center Complex Proteins/chemistry , Photosynthetic Reaction Center Complex Proteins/metabolism , Photosystem II Protein Complex , Rhodopseudomonas/chemistry , Rhodopseudomonas/metabolism , Binding Sites , Buffers , Circular Dichroism , Detergents , Energy Transfer , Glucosides , Intracellular Membranes/chemistry , Intracellular Membranes/metabolism , Photosynthetic Reaction Center Complex Proteins/isolation & purification , Temperature , Time FactorsABSTRACT
A method is described for reversibly removing bacteriochlorophyll from the B800-site of the B850-850 antenna complex from Rhodobacter sphaeroides. This method uses the oligosaccharidic detergent Triton BG-10, together with an incubation at pH 5.0. Reconstitution at the B800-site has been successfully achieved for a range of modified bacteriochlorophylls. Copyright 1998 Elsevier Science B.V. All rights reserved.
ABSTRACT
The dynamics of triplet energy transfer between the primary donor and the carotenoid were measured on several photosynthetic bacterial reaction center preparations from Rhodobacter sphaeroides: (a) wild-type strain 2.4.1, (b) strain R-26.1, (c) strain R-26.1 exchanged with 13(2)-hydroxy-[Zn]-bacteriochlorophyll at the accessory bacteriochlorophyll (BChl) sites and reconstituted with spheroidene and (d) strain R-26.1 exchanged with [3-vinyl]-13(2)-hydroxy-bacteriochlorophyll at the accessory BChl sites and reconstituted with spheroidene. The rise and decay times of the primary donor and carotenoid triplet-triplet absorption signals were monitored in the visible wavelength region between 538 and 555 nm as a function of temperature from 4 to 300 K. For the samples containing carotenoids, all of the decay times correspond well to the previously observed times for spheroidene (5 +/- 2 microseconds). The rise times of the carotenoid triplets were found in all cases to be biexponential and comprised of a strongly temperature-dependent component and a temperature-independent component. From a comparison of the behavior of the carotenoid-containing samples with that from the reaction center of the carotenoidless mutant Rb. sphaeroides R-26.1, the temperature-independent component has been assigned to the buildup of the primary donor triplet state resulting from charge recombination in the reaction center. Arrhenius plots of the buildup of the carotenoid triplet states were used to determine the activation energies for triplet energy transfer from the primary donor to the carotenoid. A model for the process of triplet energy transfer that is consistent with the data suggests that the activation barrier is strongly dependent on the triplet state energy of the accessory BChl pigment, BChlB.
