Skip to main content
Log in

Energy trapping and detrapping by wild type and mutant reaction centers of purple non-sulfur bacteria

  • Regular Paper
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Time-correlated single photon counting was used to study energy trapping and detrapping kinetics at 295 K in Rhodobacter sphaeroides chromatophore membranes containing mutant reaction centers. The mutant reaction centers were expressed in a background strain of Rb. sphaeroides which contained only B880 antenna complexes and no B800-850 antenna complexes. The excited state decay times in the isolated reaction centers from these strains were previously shown to vary by roughly 15-fold, from 3.4 to 52 ps, due to differences in the charge separation rates in the different mutants (Allen and Williams (1995) J Bioenerg Biomembr 27: 275–283). In this study, measurements were also performed on wild type Rhodospirillum rubrum and Rb. sphaeroides B880 antenna-only mutant chromatophores for comparison. The emission kinetics in membranes containing mutant reaction centers was complex. The experimental data were analyzed in terms of a kinetic model that involved fast excitation migration between antenna complexes followed by reversible energy transfer to the reaction center and charge separation. Three emission time constants were identified by fitting the data to a sum of exponential decay components. They were assigned to trapping/quenching of antenna excitations by the reaction center, recombination of the P+H charge-separated state of the reaction center reforming an emitting state, and emission from uncoupled antenna pigment-protein complexes. The first varied from 60 to 160 ps, depending on the reaction center mutation; the second was 200–300 ps, and the third was about 700 ps. The observed weak linear dependence of the trapping time on the primary charge separation time, together with the known sub-picosecond exciton migration time within the antenna, supports the concept that it is energy transfer from the antenna to the reaction center, rather than charge separation, that limits the overall energy trapping time in wild type chromatophores. The component due to charge recombination reforming the excited state is minor in wild type membranes, but increases substantially in mutants due to the decreasing free energy gap between the states P* and P+H.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

PSU:

photosynthetic unit

Bchl:

bacteriochlorophyll

Bphe:

bacteriopheophytin

P:

reaction center primary electron donor

RC:

reaction center

Rb. :

Rhodobacter

Rs. :

Rhodospirillum

EDTA:

(ethylenediamine)tetraacetic acid

Tris:

tris(hydroxymethyl)aminomethane

References

  • Abdourakhmanov IA, Danielius RV and Razjivin AP (1989) Efficiency of excitation trapping by reaction centers of complex B890 from Chromatium minutissimum. FEBS Lett 245: 47–50

    Article  Google Scholar 

  • Allen JP and Williams JC (1995) Relationship between the oxidation potential of the bacteriochlorophyll dimer and electron transfer in photosynthetic reaction centers. J Bioenerg Biomembr 27: 275–283

    PubMed  Google Scholar 

  • Bakker JGC, van Grondelle R and Den Hollander WTF (1983) Trapping, loss and annihilation of excitations in a photosynthetic system. II. Experiments with the purple bacteria Rhodospirillum rubrum and Rhodopseudomonas capsulata. Biochim Biophys Acta 725: 508–518

    Google Scholar 

  • Barvik I (1993) Computer modeling of coherence effects in excitation transfer in hexagonal PSU. Acta Phys Slov 43: 114–129

    Google Scholar 

  • Beekman LMP, van Mourik F, Jones MR, Visser HM, Hunter CN and van Grondelle R (1994) Trapping in mutants of the photosynthetic purple bacterium Rhodobacter sphaeroides: Influence of the charge separation rate and consequences for the rate-limiting step in the light-harvesting process. Biochemistry 33: 3143–3147

    PubMed  Google Scholar 

  • Bergström H, Westerhuis WHJ, Sundström V, van Grondelle R, Niederman RA and Gillbro T (1988) Energy transfer within the isolated B875 light-harvesting pigment-protein complex of Rhodobacter sphaeroides at 77 K studied by picosecond absorption spectroscopy. FEBS Lett 233: 12–16

    Article  Google Scholar 

  • Bixon M, Jortner J and Michel-Beyerle M (1995) A kinetic analysis of the primary charge separation in bacterial photosynthesis. Energy gaps and static heterogeneity. Chem Phys 197: 389–404

    Article  Google Scholar 

  • Borisov AY, Freiberg A, Godik VI, Rebane K and Timpmann K (1985) Kinetics of picosecond bacteriochlorophyll luminescence in vivo as a function of the reaction center state. Biochim Biophys Acta 807: 221–229

    Google Scholar 

  • Bradforth SE, Jimenez R, van Mourik F, van Grondelle R and Fleming GR (1995) Excitation transfer in the core light-harvesting complex (LH-1) of Rhodobacter sphaeroides: An ultrafast fluorescence depolarization and annihilation study. J Phys Chem 99: 16179–16191

    Google Scholar 

  • Deinum G, Kramer H, Aartsma TJ, Kleinherenbrink FAM and Amesz J (1991) Fluorescence quenching in Heliobacterium chlorum by reaction centers in the charge separated state. Biochim Biophys Acta 1058: 339–344

    Google Scholar 

  • Deinum G, Kleinherenbrink FAM, Aartsma TJ and Amesz J (1992) The fluorescence yield of Rhodopseudomonas viridis in relation to the redox state of the primary electron donor. Biochim Biophys Acta 1099: 81–84

    PubMed  Google Scholar 

  • Duysens LMN (1952) Transfer of excitation energy in photosynthesis. Thesis, State University of Utrecht

  • Emerson R and Arnold WA (1932) The photochemical reaction in photosynthesis. J Gen Physiol 16: 191–205

    Article  Google Scholar 

  • Feher G, Allen JP, Okamura MY and Rees DC (1989) Structure and function of bacterial photosynthetic reaction centres. Nature 339: 111–116

    Article  Google Scholar 

  • Freiberg A (1995) Coupling of antennas to reaction centers. In: Blankenship RE, Madigan MT and Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp 385–398. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Freiberg A and Timpmann K (1992) Picosecond fluorescence spectroscopy of light-harvesting antenna complexes from Rhodospirillum rubrum in the 300–4 K temperature range. Comparison with the data on chromatophores. J Photochem Photobiol B: Biol 15: 151–158

    Article  Google Scholar 

  • Freiberg A, Godik VI and Timpmann K (1984) Excitation energy transfer in bacterial photosynthesis studied by picosecond laser spectrochronography. In: Sybesma C (ed) Advances in Photosynthesis Research, Vol 1, pp 45–48. Martinus Nijhoff/Dr. W. Junk Publishers, Hague

    Google Scholar 

  • Freiberg A, Allen JP, Williams JC and Woodbury NW (1995) Trapping in mutant photosynthetic bacteria. In: Mathis P (ed) Photosynthesis: From Light to Biosphere, Vol 1, pp 863–866. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Gust D, Moore TA, Luttrull DK, Seely GR, Bittersmann E, Bensasson RV, Rougee M, Land EJ, De Schryver FC and Van der Auweraer M (1990) Photophysical properties of 2-nitro-5,10,15,20-tetra-p-tolylporphyrins. Photochem Photobiol 51: 419–426

    PubMed  Google Scholar 

  • Hunter CN (1995) Genetic manipulation of the antenna complexes of purple bacteria. In: Blankenship RE, Madigan MT and Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp 473–501. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Hunter CN, Bergström H, van Grondelle R and Sundström V (1990) Energy-transfer dynamics in three light-harvesting mutants of Rhodobacter sphaeroides: A picosecond spectroscopy study. Biochemistry 29: 3203–3207

    PubMed  Google Scholar 

  • Källebring B and Hansson Ö (1991) A theoretical study of the effect of charge recombination on the transfer and trapping of excitation energy in photosynthesis. Chem Phys 149: 361–372

    Article  Google Scholar 

  • Kirmaier C and Holten D (1987) Primary photochemistry of reaction centers from the photosynthetic purple bacteria. Photosynth Res 13: 225–260

    Google Scholar 

  • Kirmaier C and Holten D (1993) Electron transfer and charge recombination reactions in wild-type and mutant bacterial reaction centers. In: Deisenhofer J and Norris JR (eds) The Photosynthetic Reaction Center, pp 49–70. Academic Press, San Diego

    Google Scholar 

  • Kleinherenbrink FAM, Deinum G, Otte SCM, Hoff AJ and Amesz J (1992) Energy transfer from long-wavelength absorbing antenna bacteriochlorophylls to the reaction center. Biochim Biophys Acta 1099: 175–181

    Google Scholar 

  • Lin X, Williams JC, Allen JP, and Mathis P (1994) Relationship between rate and free energy difference for electron transfer from cytochrome c 2 to the reaction center in Rhodobacter sphaeroides. Biochemistry 33: 13517–13523

    PubMed  Google Scholar 

  • Martin J-L and Vos MH (1992) Femtosecond biology. Ann Rev Biophys Biomol Struct 21: 199–222

    Article  Google Scholar 

  • Mattioli TA, Lin X, Allen JP and Williams JC (1995) Correlation between multiple hydrogen bonding and alteration of the oxidation potential of the bacteriochlorophyll dimer of reaction centers from Rhodobacter sphaeroides. Biochemistry 34: 6142–6152

    PubMed  Google Scholar 

  • Müller MG, Drews G and Holzwarth AR (1993) Excitation transfer and charge separation kinetics in purple bacteria. (1) Picosecond fluorescence of chromatophores from Rhodobacter capsulatus wild type. Biochim Biophys Acta 1142: 49–58

    Google Scholar 

  • Murchison HA, Alden RG, Allen JP, Peloquin JM, Taguchi AKW, Woodbury NW and Williams JC (1993) Mutations designed to modify the environment of the primary electron donor of the reaction center from Rhodobacter sphaeroides: Phenylalanine to leucine at L167 and histidine to phenylalanine at L168. Biochemistry 32: 3498–3505

    PubMed  Google Scholar 

  • Novoderezhkin VI and Razjivin AP (1994) Exciton states of the antenna and energy trapping by the reaction center. Photosynth Res 42: 9–15

    Google Scholar 

  • Paddock ML, Rongey SH, Feher G and Okamura MY (1989) Pathway of proton transfer in bacterial reaction centers: Replacement of glutamic acid 212 in the L subunit by glutamine inhibits quinone (secondary acceptor) turnover. Proc Natl Acad Sci USA 86: 6602–6606

    PubMed  Google Scholar 

  • Parson WW (1991) Reaction centers. In: Scheer H (ed) Chlorophylls, pp 1153–1180. CRC Press, Boca Raton

    Google Scholar 

  • Peloquin JM, Williams JC, Lin X, Alden RG, Taguchi AKW, Allen JP and Woodbury NW (1994) Time-dependent thermodynamics during early electron transfer in reaction centers from Rhodobacter sphaeroides. Biochemistry 33: 8089–8100

    Google Scholar 

  • Pullerits T, Chachisvilis M and Sundström V (1995) Exciton dynamics and delocalization length in B850 molecules of LH2 of Rhodobacter sphaeroides. In: Mathis P (ed) Photosynthesis: From Light to Biosphere, Vol 1, pp 107–110. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Sebban P and Moya I (1983) Fluorescence lifetime spectra of in vivo bacteriochlorophyll at room temperature. Biochim Biophys Acta 722: 436–442

    Google Scholar 

  • Sebban P, Robert B and Jolchine G (1985) Isolation and spectroscopic characterization of the B875 antenna complex of a mutant of Rhodopseudomonas sphaeroides. Photochem Photobiol 42: 573–578

    Google Scholar 

  • Somsen OJG, Valkunas L and van Grondelle R (1996) A perturbed two-level model for excition trapping in small photosynthetic systems. Biophys J 70: 669–683

    PubMed  Google Scholar 

  • Szöcs V and Barvik I (1986) Description of excitation transfer and damping in the primary processes of bio-(chemical) systems. J Theor Biol 122: 179–186

    Google Scholar 

  • Taguchi AKW, Stocker JW, Alden RG, Causgrove TP, Peloquin JM, Boxer SG and Woodbury NW (1992) Biochemical characterization and electron-transfer reactions of syml, a Rhodobacter capsulatus reaction center symmetry mutant which affects the initial electron donor. Biochemistry 31: 10345–10355

    PubMed  Google Scholar 

  • Timpmann K, Zhang FG, Freiberg A and Sundström V (1993) Detrapping of excitation energy from the reaction centre in the photosynthetic purple bacterium Rhodospirillium rubrum. Biochim Biophys Acta 1183: 185–193

    Google Scholar 

  • Timpmann K, Freiberg A and Sundström V (1995) Energy trapping and detrapping in photosynthetic bacteria Rhodopseudomonas viridis. Chem Phys 194: 275–283

    Article  Google Scholar 

  • Valkunas L, van Mourik F and van Grondelle R (1992) On the role of spectral and spatial antenna inhomogeneity in the process of excitation energy trapping in photosynthesis. J Photochem Photobiol B: Biol 15: 159–170

    Article  Google Scholar 

  • Valkunas L, Trinkunas G, Liuolia V and van Grondelle R (1995) Nonlinear annihilation of excitations in photosynthetic systems. Biophys J 69: 1117–1129

    PubMed  Google Scholar 

  • van Grondelle R and Sundström V (1988) Excitation energy transfer in photosynthesis. In: Scheer H and Schneider S (eds) Photosynthetic Light-Harvesting Systems: Organization and Function, pp 403–438. Walter de Gruyter, Berlin, New York

    Google Scholar 

  • Visscher KJ, Bergstrom H, Sundstrom V, Hunter CN and van Grondelle R (1989) Temperature dependence of energy transfer from the long wavelength antenna BChl-896 to the reaction center in Rhodospirillum rubrum, Rhodobacter sphaeroides (w.t. and M21 mutant) from 77 to 177 K, studied by picosecond absorption spectroscopy. Photosynth Res 22: 211–217

    Google Scholar 

  • Visser HM, Somsen OJG, van Mourik F, Lin S, van Stokkum IHM and van Grondelle R (1995) Direct observation of sub-picosecond equilibration of excitation energy in the light-harvesting antenna of Rhodospirillum rubrum. Biophys J 69: 1083–1099

    PubMed  Google Scholar 

  • Vredenberg WJ and Duysens LMN (1963) Transfer of energy from bacteriochlorophyll to a reaction centre during bacterial photosynthesis. Nature 4865: 355–357

    Google Scholar 

  • Wang RT and Clayton RK (1971) The absolute yield of bacteriochlorophyll fluorescence in vivo. Photochem Photobiol 13: 215–224

    PubMed  Google Scholar 

  • Wang S, Lin X, Woodbury NW and Allen JP (1994) Comparative study of reaction centers from purple photosynthetic bateria: Isolation and optical spectroscopy. Photosynth Res 42: 203–215

    Google Scholar 

  • Williams JC and Taguchi AKW (1995) Genetic manipulation of purple photosynthetic bacteria. In: Blankenship RE, Madigan MT and Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp 1029–1065. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Williams JC, Alden RG, Murchison HA, Peloquin JM, Woodbury NW and Allen JP (1992) Effects of mutations near the bacteriochlorophylls in reaction centers from Rhodobacter sphaeroides. Biochemistry 31: 11029–11037

    PubMed  Google Scholar 

  • Woodbury NW and Allen JP (1995) The pathway, kinetics and thermodynamics of electron transfer in wild-type and mutant reaction centers of purple nonsulfur bacteria. In: Blankenship RE, Madigan MT and Bauer CE (eds) Anoxygenic Photosynthetic Bacteria, pp 527–557. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Woodbury N and Bittersmann E (1990) Time-resolved measurements of fluorescence from the photosynthetic membranes of Rhodobacter capsulatus and Rhodospirillum rubrum. In: Baltscheffsky M (ed) Current Research in Photosynthesis, Vol 2, pp 165–168. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Woodbury NW, Peloquin JM, Alden RG, Lin X, Lin S, Taguchi AKT, Williams JC and Allen JP (1994) Relationship between thermodynamics and mechanism during photoinduced charge separation in reaction centers from Rhodobacter sphaeroides. Biochemistry 33: 8101–8112

    PubMed  Google Scholar 

  • Woodbury NW, Lin S, Lin X, Peloquin JM, Taguchi AKT, Williams JC and Allen JP (1995) The role of reaction center excited state evolution during charge separation in a Rb. sphaeroides mutant with an initial electron donor midpoint potential 260 mV above wild type. Chem Phys 197: 405–421

    Article  Google Scholar 

  • Xiao W, Lin S, Taguchi AKW and Woodbury NW (1994) Femtosecond pump-probe analyses of energy and electron transfer in photosynthetic membranes of Rhodobacter capsulatus. Biochemistry 33: 8313–8322

    PubMed  Google Scholar 

  • Zinth W and Kaiser W (1993) Time-resolved spectroscopy of the primary electron transfer in reaction centers of Rhodobacter sphaeroides and Rhodopseudomonas viridis. In: Deisenhofer J and Norris JR (eds) The Photosynthetic Reaction Center, pp 71–88. Academic Press, San Diego

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Author for correspondence

Rights and permissions

Reprints and permissions

About this article

Cite this article

Freiberg, A., Allen, J.P., Williams, J.C. et al. Energy trapping and detrapping by wild type and mutant reaction centers of purple non-sulfur bacteria. Photosynth Res 48, 309–319 (1996). https://doi.org/10.1007/BF00041022

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00041022

Key words

Navigation