Skip to main content
SpringerLink
Log in

Time-resolved step-scan FTIR investigation on the primary donor of the reaction center from the green sulfur bacterium Chlorobium tepidum

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

The vibrational properties of the primary donor P840 in the reaction center (RC) of the green sulfur bacterium Chlorobium tepidum and its interactions with the surrounding protein environment have been investigated by Fourier transform infrared (FTIR) difference spectroscopy at cryogenic temperatures. By using the step-scan technique with a time resolution of 5 μs on RCs that had been depleted of the iron–sulfur electron acceptors, the formation and decay of the triplet state 3P840 have been followed in infrared for the first time. The 3P840/P840 FTIR difference spectrum is compared to the P840 +/P840 FTIR difference spectrum measured under identical conditions on untreated RCs and recorded with the same step-scan set-up. The latter P840 +/P840 difference spectrum is essentially the same as those measured under steady-state conditions using the more conventional continuous illumination method. Comparison of the 3P840/P840 and P840 +/P840 spectra provides unambiguous assignment of the vibration of the 9-keto C=O group(s) of P840 at 1684 cm−1 as the only common negative band in the two spectra. This frequency corresponds to carbonyl group(s) free from hydrogen bonding interactions. The obtained results are discussed in the framework of the structure and photochemistry of the primary donor P840.

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

Similar content being viewed by others

References

  • Breton J (2001) Fourier transform infrared spectroscopy of primary electron donors in type I photosynthetic reaction centers. Biochim Biophys Acta 1507: 180–193

    Article  PubMed  CAS  Google Scholar 

  • Breton J and Nabedryk E (1993a) S0 ? T1 infrared difference spectrum of the triplet state of the primary electron donor in Rb. sphaeroides photosynthetic bacterial reaction centers. Chem Phys Lett 213: 571–575

    Article  CAS  Google Scholar 

  • Breton J and Nabedryk E (1993b) FTIR difference spectrum of the triplet state of the primary electron donor in photosynthetic bacterial reaction centers. In: Theophanides T, Anastassopoulou J and Fotopoulos N (eds) Fifth International Conference on the Spectroscopy of Biological Molecules, pp 309–310. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Breton J, Nabedryk E and Parson WW (1992) A new infrared electronic transition of the oxidized primary donor in bacterial reaction centers: a way to assess resonance interactions between the bacteriochlorophylls. Biochemistry 31: 7503–7510

    Article  PubMed  CAS  Google Scholar 

  • Breton J, Hienerwadel R and Nabedryk E (1997) FTIR difference spectrum of the photooxidation of the primary electron donor of Photosystem II. In: Carmona P, Navarro R and Hernanz A (eds) Spectroscopy of Biological Molecules: Modern Trends, pp 101– 102. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Breton J, Nabedryk E and Leibl W (1999) FTIR Study of the primary electron donor of Photosystem I (P700) revealing delocalization of the charge in P700+ and localization of the triplet character in 3P700. Biochemistry 38: 11585–11592

    Article  PubMed  CAS  Google Scholar 

  • Brudler R and Gerwert K (1998) Step-scan FTIR spectroscopy resolves the QA-QB?QAQB-transition in Rb. sphaeroides R26reaction centers. Photosynth Res 55: 261–266

    Article  CAS  Google Scholar 

  • Burie J-R, Leibl W, Nabedryk E and Breton J (1993) Step-scan FT-IR spectroscopy of electron transfer in the photosynthetic bacterial reaction center. Appl Spectr 47: 1401–1404

    Article  CAS  Google Scholar 

  • El-Kabbani O, Chang C-H, Tiede D, Norris J and Schiffer M (1991) Comparison of reaction centers from Rhodobacter sphaeroides and Rhodopseudomonas viridis: overall architecture and protein– pigment interactions. Biochemistry 30: 5361–5369.

    Article  PubMed  CAS  Google Scholar 

  • Gerwert K (2000) Time-resolved FT-IR difference spectroscopy: a tool to monitor molecular reaction mechanisms of proteins. In: Gremilch H-U and Yan B (eds) Infrared and Raman Spectroscopy of Biological Materials, Vol 2, pp 193–230. Marcel Dekker, New York

    Google Scholar 

  • Guergova-Kuras M, Boudreaux B, Joliot A, Joliot P and Redding K (2001) Evidence for two active branches for electron transfer in Photosystem I. Proc Natl Acad Sci USA 98: 4437–4442

    CAS  Google Scholar 

  • Hastings G (2001) Time-resolved step-scan Fourier transform infrared and visible absorption difference spectroscopy for the study of Photosystem I. Appl Spectr 55: 894–900

    Article  CAS  Google Scholar 

  • Hauska G, Schoedl T, Remigy H and Tsiotis G (2001) The reaction center of green sulfur bacteria. Biochim Biophys Acta 1507: 260–277.

    Article  PubMed  CAS  Google Scholar 

  • Iwaki M, Itoh S, Kamei S, Matsubara H and Oh-oka H (1999) Timeresolved spectroscopy of Chlorophyll-a like electron acceptor in the reaction center complex of the green sulfur bacterium Chlorobium tepidum. Plant Cell Physiol 40: 1021–1028

    CAS  Google Scholar 

  • Jordan P, Fromme P, Witt HT, Saenger W and Krauß N (2001) Three-dimensional structure of cyanobacterial Photosystem I at 2.5°A resolution. Nature 411: 909–917

    Article  PubMed  CAS  Google Scholar 

  • Kusumoto N, Sétif P, Brettel K, Seo D and Sakurai H (1999) Electron transfer kinetics in purified reaction centers from the green sulfur bacterium Chlorobium tepidum studied by multiple-flash excitation. Biochemistry 38: 12124–12137

    Article  PubMed  CAS  Google Scholar 

  • Mäntele W, Wollenweber AM, Nabedryk E and Breton J (1988) Infrared spectroelectrochemistry of bacteriochlorophylls and bacteriopheophytins: implication for the binding of the pigments in the reaction center from photosynthetic bacteria. Proc Natl Acad Sci USA 85: 8468–8472

    Article  PubMed  Google Scholar 

  • Nabedryk E (1996) Light-induced Fourier trasform infrared difference spectroscopy of the primary electron donor in photosynthetic reaction centers. In: Mantsch HH and Chapman D (eds) Infrared Spectroscopy of Biomolecules, pp 39–81. Wiley-Liss New York

    Google Scholar 

  • Nabedryk E, Allen JP, Taguchi AKW, Williams JC, Woodbury NW and Breton J (1993) Fourier transform infrared study of the primary electron donor in chromatophores of Rhodobacter sphaeroides with reaction centers genetically modified at residues M160 and L131. Biochemistry 32: 13879–13885

    Article  PubMed  CAS  Google Scholar 

  • Nabedryk E, Leibl W and Breton J (1996) FTIR spectroscopy of primary donor photooxidation in Photosystem I, Heliobacillus mobilis, and Chlorobium limicola. Comparison with purple bacteria. Photosynth Res 48: 301–308

    Article  CAS  Google Scholar 

  • Noguchi T, Inoue Y and Satoh K (1993) FT-IR studies on the triplet state of P680 in the Photosystem II reaction center: triplet equilibrium within a chlorophyll dimer. Biochemistry 32: 7186–7195

    Article  PubMed  CAS  Google Scholar 

  • Noguchi T, Kusumoto N, Inoue Y and Sakurai H (1996) Electronic and vibrational structure of the radical cation of P840 in the putative reaction center from Chlorobium tepidum as studied by FTIR spectroscopy. Biochemistry 35: 15428–15435

    Article  PubMed  CAS  Google Scholar 

  • Noguchi T, Tomo T and Inoue Y (1998) Fourier transform infrared study of the cation radical of P680 in the Photosystem II reaction center: evidence for charge delocalisation on the chlorophyll dimer. Biochemistry 37: 13164–13625

    Article  Google Scholar 

  • Reimers JR and Hush NS (1995) Nature of the ground and first excited states of the radical cations of photosynthetic bacterial reaction centers. Chem Phys 197: 323–332

    Article  CAS  Google Scholar 

  • Rödig C and Siebert F (1999) Error and artefacts in time-resolved step-scan FT-IR spectroscopy. Appl Spectr 53: 893–901

    Article  Google Scholar 

  • Sakurai H, Kusumoto N and Inoue K (1996) Function of the reaction center of green sulfur bacteria. Photochem Photobiol 64: 5–13

    CAS  Google Scholar 

  • Scott MP, Kjoer B, Scheller HV and Golbeck JH (1997) Redox titration of two [4Fe–4S] clusters in the photosynthetic reaction center from the anaerobic green sulfur bacterium Chlorobium vibrioforme. Eur J Biochem 244: 454–461

    Article  PubMed  CAS  Google Scholar 

  • Seo D and Sakurai H (2002) Purification and characterization of ferrodoxin-NAD(P)+ reductase from the green sulfur bacterium Chlorobium tepidum. Biochim Biophys Acta – Protein Struct Mol Enzymol 1597: 123–132

    Article  CAS  Google Scholar 

  • Seo D, Tomioka A, Kusumoto N, Kamo M, Enami I and Sakurai H (2001) Purification of ferrodoxins and their reaction with purified reaction center complex from the green sulfur bacterium Chlorobium tepidum. Biochim Biophys Acta – Bioenergetics 1503: 377–384

    Article  CAS  Google Scholar 

  • Sétif P, Seo D and Sakurai H (2001) Photoreduction and reoxidation of the three iron sulfur clusters of reaction centers of green sulfur bacteria. Biophys J 81: 1208–1219

    Article  PubMed  Google Scholar 

  • Thibodeau DL, Nabedryk E and Breton J (1991) Light-induced polarized FTIR spectroscopy of oriented photosynthetic bacterial reaction centers. In: Michel-Beyerle ME (ed) Reaction Centers of Photosynthetic Bacteria. Series in Biophysics, Vol 6, pp 87–98. Springer-Verlag, Berlin

    Google Scholar 

  • Uhmann W, Becker A, Taran C and Siebert F (1991) Time-Resolved FT-IR absorption spectroscopy using a step-scan interferometer. Appl Spectr 45: 390–397

    Article  CAS  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, The Netherlands

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Service de Bioénergétique, CEA-Saclay, Bât. 532, 91191, Gif-sur-Yvette cedex, France

    Alberto Mezzetti, Winfried Leibl & Jacques Breton

  2. Department of Physical Chemistry `A. Miolati', University of Padova, Via Loredan 2, 35100, Padova, Italy

    Alberto Mezzetti

  3. Department of Biology, School of Education, Waseda University, Nishiwaseda, Shinjuku, Tokyo, 169-8050, Japan

    Daisuke Seo & Hidehiro Sakurai

Authors
  1. Alberto Mezzetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Daisuke Seo
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Winfried Leibl
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hidehiro Sakurai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jacques Breton
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mezzetti, A., Seo, D., Leibl, W. et al. Time-resolved step-scan FTIR investigation on the primary donor of the reaction center from the green sulfur bacterium Chlorobium tepidum . Photosynthesis Research 75, 161–169 (2003). https://doi.org/10.1023/A:1022867317267

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1022867317267

Search

Navigation