title: Function of three cytochromes in photosynthesis of whole cells of Rhodospirillum rubrum as studied by flash spectroscopy. Evidence for two types of reaction center.
abstract: 1. Changes in the absorption spectrum induced by 10-mus flashes and continuous light of various intensities were studied in whole cells of Rhodospirillum rubrum in the presence and absence of 2-n-heptyl-4-hydroxyquinoline-N-oxide(HOQNO) and antimycin A. 2. Three cytochromes, c-420 (cytochrome c2), c-560 (cytochrome b) and c-428 were photoactive and gamma and alpha peaks at 420 and 550, 428 and 560, and 428 and 551 nm, respectively; they were photooxidized following the flash with half times of 0.3, 0.6 and 7 ms in the approximate ratios of 1/100, 1/300 and 1/1000 (cytochrome oxidized/antenna chlorophyll) and became reduced with half times of 12 ms, 60 ms and 0.7 s, respectively. c-428 and c-560 have not been distinguished before. 3. From a detailed analysis of the kinetics of P+ (oxidized reaction center chlorophyll) and the cytochromes, we conclude that 5% of the P+ (P2+) oxidizes c-428, whereas the remaining 95% of P+ (P1+) oxidizes c-420. At actinic light intensities low enough to keep c-420 fully reduced, approx. 4-5% of P becomes oxidized, accompanied by all c-428. The P2+ -P2 difference spectrum induced by this weak light is, when corrected for a shift to longer wavelengths of the bacteriochlorophyll absorption band at 878 nm, identical to the difference spectrum caused by the photooxidation of the remaining P1. At low flash intensity, c-428 becomes preferentially photooxidized, which suggests that the reaction centers where c-428 functions as a secondary donor contain much more antenna pigments compared to the centers where c-420 serves this purpose. 4. c+-420 is reduced in a competitive way by reduced c-560 (t 1/2=7 ms), and by an electron donor pool, (t 1/2=15 ms). HOQNO inhibits both pathways; antimycin A only the first. In the presence of HOQNO, c-560 is in the oxidized state in the dark, and is reduced in a light flash (t 1/2=100 ms), indicating that c-560 acts in a cyclic electron transport chain connected to P1. 5. The ratio of numbers of molecules P1 and antenna bacteriochlorophyll, transferring excitation energy to P1, is P1/bacteriochlorophyll1=1/30 P2: bacteriochlorophyll2=1/300; c-420/P1=1:2; c-560/P1=1/6; C-428/P2=1/1; bacteriochlorophyll2=7:3. If P2 is oxidized, excitation energy is transferred from bacteriochlorophyll2 to bacteriochlorophyll1.
mesh_terms: D000693:Anaerobiosis; D002734:Chlorophyll / Q000378:metabolism; D003201:Computers; D003574:Cytochrome c Group / Q000378:metabolism; D003580:Cytochromes / Q000378:metabolism*; D007700:Kinetics; D007834:Lasers; D008027:Light; D010788:Photosynthesis; D012247:Rhodospirillum rubrum / Q000201:enzymology*; D013053:Spectrophotometry; D013057:Spectrum Analysis
chemical_list: D003574:Cytochrome c Group; D003580:Cytochromes; D002734:Chlorophyll