Skip to main content

Advertisement

SpringerLink
Log in

Ultrafast time-resolved spectroscopy of the light-harvesting complex 2 (LH2) from the photosynthetic bacterium Thermochromatium tepidum

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

Abstract

The light-harvesting complex 2 from the thermophilic purple bacterium Thermochromatium tepidum was purified and studied by steady-state absorption and fluorescence, sub-nanosecond-time-resolved fluorescence and femtosecond time-resolved transient absorption spectroscopy. The measurements were performed at room temperature and at 10 K. The combination of both ultrafast and steady-state optical spectroscopy methods at ambient and cryogenic temperatures allowed the detailed study of carotenoid (Car)-to-bacteriochlorophyll (BChl) as well BChl-to-BChl excitation energy transfer in the complex. The studies show that the dominant Cars rhodopin (N = 11) and spirilloxanthin (N = 13) do not play a significant role as supportive energy donors for BChl a. This is related with their photophysical properties regulated by long π-electron conjugation. On the other hand, such properties favor some of the Cars, particularly spirilloxanthin (N = 13) to play the role of the direct quencher of the excited singlet state of BChl.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9

Similar content being viewed by others

Abbreviations

BChl:

Bacteriochlorophyll a

LH2:

Light-harvesting complex 2

LH1-RC:

Light-harvesting complex 1-reaction center

ICM:

Intracytoplasmic membrane

DDM:

N-Dodecyl β-d-maltoside

TCSPC:

Time-correlated single photon counting

EET:

Efficiency of energy transfer

RT:

Room temperature

Car:

Carotenoid

VIS:

Visible

NIR:

Near-infrared

ESA:

Excited state absorption

EADS:

Evolution-associated difference spectrum

IRF:

Instrument response function

References

  • Andersson PO, Gillbro T (1995) Photophysics and dynamics of the lowest excited singlet state in long substituted polyenes with implications to the very long chain limit. J Chem Phys 103:2509–2519

    Article  CAS  Google Scholar 

  • Billsten HH, Herek JL, Garcia-Asua G, Hashoj L, Polivka T, Hunter CN, Sundstrom V (2002) Dynamics of energy transfer from lycopene to bacteriochlorophyll in genetically-modified LH2 complexes of Rhodobacter sphaeroides. Biochemistry 41:4127–4136

    Article  CAS  Google Scholar 

  • Billsten HH, Bhosale P, Yemelyanov A, Bernstein PS, Polivka T (2003) Photophysical properties of xanthophylls in carotenoproteins from human retinas. Photochem Photobiol 78:138–145

    Article  PubMed  CAS  Google Scholar 

  • Billsten HH, Pan J, Sinha S, Pascher T, Sundstrom V, Polivka T (2005) Excited-state processes in the carotenoid zeaxanthin after excess energy excitation. J Phys Chem A 109:6852–6859

    Article  PubMed  CAS  Google Scholar 

  • Bopp MA, Jia YW, Li LQ, Cogdell RJ, Hochstrasser RM (1997) Fluorescence and photobleaching dynamics of single light-harvesting complexes. Proc Natl Acad Sci USA 94:10630–10635

    Article  PubMed  CAS  Google Scholar 

  • Buckup T, Savolainen J, Wohlleben W, Herek JL, Hashimoto H, Correia RR, Motzkus M (2006) Pump-probe and pump-deplete-probe spectroscopies on carotenoids with N = 9–15 conjugated bonds. J Chem Phys 125:194505

    Article  PubMed  Google Scholar 

  • Chen XH, Zhang L, Weng YX, Du LC, Ye MP, Yang GZ, Fujii R, Rondonuwu FS, Koyama Y, Wu YS, Zhang JP (2005) Protein structural deformation induced lifetime shortening of photosynthetic bacteria light-harvesting complex LH2 excited state. Biophys J 88:4262–4273

    Article  PubMed  CAS  Google Scholar 

  • Cogdell RJ, Scheer H (1985) Circular dichroism of light harvesting complexes from purple photosynthetic bacteria. Photochem Photobiol 42:669–678

    Article  CAS  Google Scholar 

  • Cong H, Niedzwiedzki DM, Gibson GN, LaFountain AM, Kelsh RM, Gardiner AT, Cogdell RJ, Frank HA (2008) Ultrafast time-resolved carotenoid to-bacteriochlorophyll energy transfer in LH2 complexes from photosynthetic bacteria. J Phys Chem B 112:10689–10703

    Article  PubMed  CAS  Google Scholar 

  • Connolly JS, Samuel EB, Janzen AF (1982) Effects of solvent on the fluorescence properties of bacteriochlorophyll a. Photochem Photobiol 36:565–574

    Article  CAS  Google Scholar 

  • Fiedor L, Akahane J, Koyama Y (2004) Carotenoid-induced cooperative formation of bacterial photosynthetic LH1 complex. Biochemistry 43:16487–16496

    Article  PubMed  CAS  Google Scholar 

  • Frank HA, Desamero RZB, Chynwat V, Gebhard R, van der Hoef I, Jansen FJ, Lugtenburg J, Gosztola D, Wasielewski MR (1997) Spectroscopic properties of spheroidene analogs having different extents of π-electron conjugation. J Phys Chem A 101:149–157

    Article  CAS  Google Scholar 

  • Freiberg A, Ratsep M, Timpmann K, Trinkunas G, Woodbury NW (2003) Self-trapped excitons in LH2 antenna complexes between 5 K and ambient temperature. J Phys Chem B 107:11510–11519

    Article  CAS  Google Scholar 

  • Gradinaru CC, Kennis JT, Papagiannakis E, van Stokkum IH, Cogdell RJ, Fleming GR, Niederman RA, van Grondelle R (2001) An unusual pathway of excitation energy deactivation in carotenoids: singlet-to-triplet conversion on an ultrafast timescale in a photosynthetic antenna. Proc Natl Acad Sci USA 98:2364–2369

    Article  PubMed  CAS  Google Scholar 

  • Hayashi H, Nozawa T, Hatano M, Morita S (1981) Circular dichroism of bacteriochlorophyll a in light harvesting bacteriochlorophyll protein complexes from Chromatium vinosum. J Biochem 89:1853–1861

    PubMed  CAS  Google Scholar 

  • Jailaubekov AE, Song SH, Vengris M, Cogdell RJ, Larsen DS (2010) Using narrowband excitation to confirm that the S* state in carotenoids is not a vibrationally-excited ground state species. Chem Phys Lett 487:101–107

    Article  CAS  Google Scholar 

  • Kennis JTM, Streltsov AM, Aartsma TJ, Nozawa T, Amesz J (1996) Energy transfer and exciton coupling in isolated B800-850 complexes of the photosynthetic purple sulfur bacterium Chromatium tepidum. The effect of structural symmetry on bacteriochlorophyll excited states. J Phys Chem 100:2438–2442

    Article  CAS  Google Scholar 

  • Kereiche S, Bourinet L, Keegstra W, Arteni AA, Verbavatz JM, Boekema EJ, Robert B, Gall A (2008) The peripheral light-harvesting complexes from purple sulfur bacteria have different ‘ring’ sizes. FEBS Lett 582:3650–3656

    Article  PubMed  CAS  Google Scholar 

  • Kobayashi M, Fujioka Y, Mori T, Terashima M, Suzuki H, Shimada Y, Saito T, Wang ZY, Nozawa T (2005) Reconstitution of photosynthetic reaction centers and core antenna-reaction center complexes in liposomes and their thermal stability. Biosci Biotechnol Biochem 69:1130–1136

    Article  PubMed  CAS  Google Scholar 

  • Larsen DS, Papagiannakis E, van Stokkum IHM, Vengris M, Kennis JTM, van Grondelle R (2003) Excited state dynamics of beta-carotene explored with dispersed multi-pulse transient absorption. Chem Phys Lett 381:733–742

    Article  CAS  Google Scholar 

  • Ma F, Kimura Y, Zhao XH, Wu YS, Wang P, Fu LM, Wang ZY, Zhang JP (2008) Excitation dynamics of two spectral forms of the core complexes from photosynthetic bacterium Thermochromatium tepidum. Biophys J 95:3349–3357

    Article  PubMed  CAS  Google Scholar 

  • Ma F, Kimura Y, Yu LJ, Wang P, Ai XC, Wang ZY, Zhang JP (2009) Specific Ca2+-binding motif in the LH1 complex from photosynthetic bacterium Thermochromatium tepidum as revealed by optical spectroscopy and structural modeling. FEBS J 276:1739–1749

    Article  PubMed  CAS  Google Scholar 

  • Macpherson AN, Gillbro T (1998) Solvent dependence of the ultrafast S2-S1 internal conversion rate of β-carotene. J Phys Chem A 102:5049–5058

    Article  CAS  Google Scholar 

  • Macpherson AN, Arellano JB, Fraser NJ, Cogdell RJ, Gillbro T (2001) Efficient energy transfer from the carotenoid S2 state in a photosynthetic light-harvesting complex. Biophys J 80:923–930

    Article  PubMed  CAS  Google Scholar 

  • Madigan MT (1984) A novel photosynthetic purple bacterium isolated from a Yellowstone hot spring. Science 225:313–315

    Article  PubMed  CAS  Google Scholar 

  • Madigan MT (1986) Chromatium Tepidum Sp-Nov, a thermophilic photosynthetic bacterium of the family Chromatiaceae. Int J Syst Bacteriol 36:222–227

    Article  CAS  Google Scholar 

  • Makhneva Z, Bolshakov M, Moskalenko A (2008) Heterogeneity of carotenoid content and composition in LH2 of the purple sulphur bacterium Allochromatium minutissimum grown under carotenoid-biosynthesis inhibition. Photosynth Res 98:633–641

    Article  PubMed  CAS  Google Scholar 

  • Monshouwer R, Abrahamsson M, van Mourik F, van Grondelle R (1997) Superradiance and exciton delocalization in bacterial photosynthetic light-harvesting systems. J Phys Chem B 101:7241–7248

    Article  CAS  Google Scholar 

  • Musewald C, Hartwich G, Pollinger-Dammer F, Lossau H, Scheer H, Michel-Beyerle ME (1998) Time-resolved spectral investigation of bacteriochlorophyll a and its transmetalated derivatives Zn-bacteriochlorophyll a and Pd-bacteriochlorophyll a. J Phys Chem B 102:8336–8342

    Article  CAS  Google Scholar 

  • Niedzwiedzki DM, Blankenship RE (2010) Singlet and triplet excited state properties of natural chlorophylls and bacteriochlorophylls. Photosynth Res 106:227–238

    Article  PubMed  CAS  Google Scholar 

  • Niedzwiedzki DM, Sullivan JO, Polivka T, Birge RR, Frank HA (2006) Femtosecond time-resolved transient absorption spectroscopy of xanthophylls. J Phys Chem B 110:22872–22885

    Article  PubMed  CAS  Google Scholar 

  • Niedzwiedzki D, Koscielecki JF, Cong H, Sullivan JO, Gibson GN, Birge RR, Frank HA (2007) Ultrafast dynamics and excited state spectra of open-chain carotenoids at room and low temperatures. J Phys Chem B 111:5984–5998

    Article  PubMed  CAS  Google Scholar 

  • Niedzwiedzki DM, Sandberg DJ, Cong H, Sandberg MN, Gibson GN, Birge RR, Frank HA (2009) Ultrafast time-resolved absorption spectroscopy of geometric isomers of carotenoids. Chem Phys 357:4–16

    Article  CAS  Google Scholar 

  • Niedzwiedzki DM, Enriquez MM, Lafountain AM, Frank HA (2010) Ultrafast time-resolved absorption spectroscopy of geometric isomers of xanthophylls. Chem Phys 373:80–89

    Article  PubMed  CAS  Google Scholar 

  • Niedzwiedzki DM, Kobayashi M, Blankenship RE (2011) Triplet excited state spectra and dynamics of carotenoids from the thermophilic purple photosynthetic bacterium Thermochromatium tepidum. Photosynth Res 107:177–186

    Article  PubMed  CAS  Google Scholar 

  • Nozawa T, Fukada T, Hatano M, Madigan MT (1986) Organization of intracytoplasmic membranes in a novel thermophilic purple photosynthetic bacterium as revealed by absorption, circular dichroism and emission spectra. Biochim Biophys Acta 852:191–197

    Article  CAS  Google Scholar 

  • Okamoto H, Ogura M, Nakabayashi T, Tasumi M (1998) Sub-picosecond excited-state dynamics of a carotenoid (spirilloxanthin) in the light-harvesting systems of Chromatium vinosum. Relaxation process from the optically allowed S2 state. Chem Phys 236:309–318

    Article  CAS  Google Scholar 

  • Papagiannakis E, Kennis JT, van Stokkum IH, Cogdell RJ, van Grondelle R (2002) An alternative carotenoid-to-bacteriochlorophyll energy transfer pathway in photosynthetic light harvesting. Proc Natl Acad Sci USA 99:6017–6022

    Article  PubMed  CAS  Google Scholar 

  • Papagiannakis E, van Stokkum IHM, van Grondelle R, Niederman RA, Zigmantas D, Sundstrom V, Polivka T (2003) A near-infrared transient absorption study of the excited-state dynamics of the carotenoid spirilloxanthin in solution and in the LH1 complex of Rhodospirillum rubrum. J Phys Chem B 107:11216–11223

    Article  CAS  Google Scholar 

  • Papagiannakis E, van Stokkum IH, Vengris M, Cogdell RJ, van Grondelle R, Larsen DS (2006) Excited-state dynamics of carotenoids in light-harvesting complexes. 1. Exploring the relationship between the S1 and S* states. J Phys Chem B 110:5727–5736

    Article  PubMed  CAS  Google Scholar 

  • Pendon ZD, Gibson GN, van der Hoef I, Lugtenburg J, Frank HA (2005) Effect of isomer geometry on the steady-state absorption spectra and femtosecond time-resolved dynamics of carotenoids. J Phys Chem B 109:21172–21179

    Article  PubMed  CAS  Google Scholar 

  • Pflock T, Dezi M, Venturoli G, Cogdell RJ, Kohler J, Oellerich S (2008) Comparison of the fluorescence kinetics of detergent-solubilized and membrane-reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides. Photosynth Res 95:291–298

    Article  PubMed  CAS  Google Scholar 

  • Polivka T, Zigmantas D, Frank HA, Bautista JA, Herek JL, Koyama Y, Fujii R, Sundstrom V (2001) Near-infrared time-resolved study of the S1 state dynamics of the carotenoid spheroidene. J Phys Chem B 105:1072–1080

    Article  CAS  Google Scholar 

  • Polivka T, Zigmantas D, Herek JL, He Z, Pascher T, Pullerits T, Cogdell RJ, Frank HA, Sundstrom V (2002) The carotenoid S1 state in LH2 complexes from purple bacteria Rhodobacter sphaeroides and Rhodopseudomonas acidophila: S1 energies, dynamics, and carotenoid radical formation. J Phys Chem B 106:11016–11025

    Article  CAS  Google Scholar 

  • Stepanenko I, Kompanetz V, Makhneva Z, Chekalin S, Moskalenko A, Razjivin A (2009) Two-photon excitation spectroscopy of carotenoid-containing and carotenoid-depleted LH2 complexes from purple bacteria. J Phys Chem B 113:11720–11723

    Article  PubMed  CAS  Google Scholar 

  • Theiss C, Leupold D, Moskalenko AA, Razjivin AP, Eichler HJ, Lokstein H (2008) Femtosecond spectroscopy of native and carotenoidless purple-bacterial LH2 clarifies functions of carotenoids. Biophys J 94:4808–4811

    Article  PubMed  CAS  Google Scholar 

  • van Dijk B, Nozawa T, Hoff AJ (1998) The B800-850 complex of the purple bacterium Chromatium tepidum: low-temperature absorption and Stark spectra. Spectrochim Acta A 54:1269–1278

    Article  Google Scholar 

  • van Stokkum IH, Larsen DS, van Grondelle R (2004) Global and target analysis of time-resolved spectra. Biochim Biophys Acta 1657:82–104

    Article  PubMed  Google Scholar 

  • Wohlleben W, Buckup T, Herek JL, Cogdell RJ, Motzkus M (2003) Multichannel carotenoid deactivation in photosynthetic light harvesting as identified by an evolutionary target analysis. Biophys J 85:442–450

    Article  PubMed  CAS  Google Scholar 

  • Wohlleben W, Buckup T, Hashimoto H, Cogdell RJ, Herek JL, Motzkus M (2004) Pump-deplete-probe spectroscopy and the puzzle of carotenoid dark states. J Phys Chem B 108:3320–3325

    Article  CAS  Google Scholar 

  • Yang F, Yu LJ, Wang P, Ai XC, Wang ZY, Zhang JP (2010) Excitation dynamics of the light-harvesting complex 2 from Thermochromatium tepidum. Acta Phys Chim Sin 26:2021–2030

    CAS  Google Scholar 

  • Yang F, Yu LJ, Wang P, Ai XC, Wang ZY, Zhang JP (2011) Effects of aggregation on the excitation dynamics of LH2 from Thermochromatium tepidum in aqueous phase and in chromatophores. J Phys Chem B 115:7906–7913

    Article  PubMed  CAS  Google Scholar 

  • Zhang JP, Fujii R, Qian P, Inaba T, Mizoguchi T, Onaka K, Watanabe Y, Nagae H, Koyama Y (2000) Mechanism of the carotenoid-to-bacteriochlorophyll energy transfer via the S1 state in the LH2 complexes from purple bacteria. J Phys Chem B 104:3683–3691

    Article  CAS  Google Scholar 

Download references

Acknowledgments

This material is based on work supported as part of the Photosynthetic Antenna Research Center (PARC), an Energy Frontier Research Center funded by the U.S. Department of Energy, Office of Science, Office of Basic Energy Sciences under Award Number DE-SC 0001035. Work in the laboratory of HAF was supported by a grant from the National Science Foundation (MCB-0913022). Author contributions: DMN-purified carotenoids, carried out the spectroscopic measurements, analyzed the data and wrote the paper; MK grew the bacterium and purified LH2, MF carried out the spectroscopic measurements; HAF and REB supervised the project.

Author information

Authors and Affiliations

  1. Photosynthetic Antenna Research Center, Washington University in St. Louis, Campus Box 1138, St. Louis, MO, 63130, USA

    Dariusz M. Niedzwiedzki & Robert E. Blankenship

  2. Department of Chemistry, University of Connecticut, U-3060, 55 North Eagleville Road, Storrs, CT, 06269-3060, USA

    Marcel Fuciman & Harry A. Frank

  3. Department of Materials Science and Chemical Engineering, Ariake National College of Technology, 150 Higashihagio-Machi, Omuta, Fukuoka, 836-85854, Japan

    Masayuki Kobayashi

  4. Department of Biology, Washington University in St. Louis, St. Louis, MO, 63130, USA

    Robert E. Blankenship

  5. Department of Chemistry, Washington University in St. Louis, St. Louis, MO, 63130, USA

    Robert E. Blankenship

Authors
  1. Dariusz M. Niedzwiedzki
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marcel Fuciman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Masayuki Kobayashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Harry A. Frank
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Robert E. Blankenship
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dariusz M. Niedzwiedzki.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Niedzwiedzki, D.M., Fuciman, M., Kobayashi, M. et al. Ultrafast time-resolved spectroscopy of the light-harvesting complex 2 (LH2) from the photosynthetic bacterium Thermochromatium tepidum . Photosynth Res 110, 49–60 (2011). https://doi.org/10.1007/s11120-011-9692-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11120-011-9692-7

Keywords

Search

Navigation