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Self-assembly and energy transfer in artificial light-harvesting complexes of bacteriochlorophyll c with astaxanthin

Abstract

Chlorosomes, the light-harvesting antennae of green photosynthetic bacteria, are based on large aggregates of bacteriochlorophyll molecules. Aggregates with similar properties to those in chlorosomes can also be prepared in vitro. Several agents were shown to induce aggregation of bacteriochlorophyll c in aqueous environments, including certain lipids, carotenes, and quinones. A key distinguishing feature of bacteriochlorophyll c aggregates, both in vitro and in chlorosomes, is a large (>60 nm) red shift of their Qy absorption band compared with that of the monomers. In this study, we investigate the self-assembly of bacteriochlorophyll c with the xanthophyll astaxanthin, which leads to the formation of a new type of complexes. Our results indicate that, due to its specific structure, astaxanthin molecules competes with bacteriochlorophylls for the bonds involved in the aggregation, thus preventing the formation of any significant red shift compared with pure bacteriochlorophyll c in aqueous buffer. A strong interaction between both the types of pigments in the developed assemblies, is manifested by a rather efficient (~40%) excitation energy transfer from astaxanthin to bacteriochlorophyll c, as revealed by fluorescence excitation spectroscopy. Results of transient absorption spectroscopy show that the energy transfer is very fast (<500 fs) and proceeds through the S2 state of astaxanthin.

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References

  1. Alster J, Zupcanova A, Vacha F, Psencik J (2008) Effect of quinones on formation and properties of bacteriochlorophyll c aggregates. Photosynth Res 95:183–189

  2. Alster J, Polivka T, Arellano JB, Chabera P, Vacha F, Psencik J (2010) beta-Carotene to bacteriochlorophyll c energy transfer in self-assembled aggregates mimicking chlorosomes. Chem Phys 373:90–97

  3. Arellano JB, Psencik J, Borrego CM, Ma YZ, Guyoneaud R, Garcia-Gil J, Gillbro T (2000) Effect of carotenoid biosynthesis inhibition on the chlorosome organization in Chlorobium phaeobacteroides strain CL1401. Photochem Photobiol 71:715–723

  4. Arellano JB, Melo TB, Fyfe PK, Cogdell RJ, Naqvi KR (2004) Multichannel flash spectroscopy of the reaction centers of wild-type and mutant Rhodobacter sphaeroides: bacteriochlorophyll B -mediated interaction between the carotenoids triplet and the special pair. Photochem Photobiol 79:68–75

  5. Balaban TS (2005) Tailoring porphyrins and chlorins for self-assembly in biomimetic artificial antenna systems. Acc Chem Res 38:612–623

  6. Balaban TS, Tamiaki H, Holzwarth AR (2005) Chlorins programmed for self-assembly. Top Curr Chem 258:1–38

  7. 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

  8. Blankenship RE, Matsuura K (2003) Antenna complexes from green photosynthetic bacteria. In: Green BR, Parson WW (eds) Light-harvesting antennas in photosynthesis. Kluwer Academic Publishers, Dordrecht, pp 195–217

  9. Bryant DA, Vassilieva EV, Frigaard NU, Li H (2002) Selective protein extraction from Chlorobium tepidum chlorosomes using detergents. Evidence that CsmA forms multimers and binds bacteriochlorophyll a. Biochemistry 41:14403–14411

  10. Chabera P, Fuciman M, Naqvi KR, Polivka T (2010) Ultrafast dynamics of hydrophilic carbonyl carotenoids—relation between structure and excited-state properties in polar solvents. Chem Phys 373:56–64

  11. Christensson N, Polivka T, Yartsev A, Pullerits T (2009) Photon echo spectroscopy reveals structure-dynamics relationships in carotenoids. Phys Rev B 79, 245118

  12. Frigaard NU, Bryant DA (2006) Chlorosomes: antenna organelles in photosynthetic green bacteria. In: Shively JM (ed) Complex intracellular structures in prokaryotes (series: Microbiology Monographs, vol. 2), pp 79–114. Springer, Berlin

  13. Frigaard NU, Takaichi S, Hirota M, Shimada K, Matsuura K (1997) Quinones in chlorosomes of green sulfur bacteria and their role in the redox-dependent fluorescence studied in chlorosome-like bacteriochlorophyll c aggregates. Arch Microbiol 167:343–349

  14. Ganapathy S, Oostergetel GT, Wawrzyniak PK, Reus M, Chew AGM, Buda F, Boekema EJ, Bryant DA, Holzwarth AR, de Groot HJM (2009) Alternating syn-anti bacteriochlorophylls form concentric helical nanotubes in chlorosomes. Proc Natl Acad Sci 106:8525–8530

  15. Gradinaru CC, Kennis JTM, Papagiannakis E, van Stokkum IHM, 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 98:2364–2369

  16. Hildebrandt P, Tamiaki H, Holzwarth AR, Schaffner K (1994) Resonance Raman-spectroscopic study of metallochlorin aggregates Implications for the supramolecular structure in chlorosomal BChl c antennae of green bacteria. J Phys Chem 98:2192–2197

  17. Hirota M, Moriyama T, Shimada K, Miller M, Olson JM, Matsuura K (1992) High degree of organization of bacteriochlorophyll c in chlorosome-like aggregates spontaneously assembled in aqueous solution. Biochim Biophys Acta 1099:271–274

  18. Ilagan RP, Christensen RL, Chapp TW, Gibson GN, Pascher T, Polivka T, Frank HA (2005) Femtosecond time-resolved absorption spectroscopy of astaxanthin in solution and in alpha-crustacyanin. J Phys Chem A 109:3120–3127

  19. Klinger P, Arellano JB, Vacha FE, Hala J, Psencik J (2004) Effect of carotenoids and monogalactosyl diglyceride on bacteriochlorophyll c aggregates in aqueous buffer: implications for the self-assembly of chlorosomes. Photochem Photobiol 80:572–578

  20. Kopczynski M, Lenzer T, Oum K, Seehusen J, Seidel MT, Ushakov VG (2005) Ultrafast transient lens spectroscopy of various C-40 carotenoids: lycopene, beta-carotene, (3R,3′ R)- zeaxanthin, (3R,3′ R,6′ R)-lutein, echinenone, canthaxanthin, and astaxanthin. Phys Chem Chem Phys 7:2793–2803

  21. Latimer P, Eubanks CAH (1962) Absorption spectrophotometry of turbid suspensions—a method of correcting for large systematic distortions. Arch Biochem Biophys 98:274–285

  22. Melo TB, Frigaard NU, Matsuura K, Naqvi KR (2000) Electronic energy transfer involving carotenoid pigments in chlorosomes of two green bacteria: Chlorobium tepidum and Chloroflexus aurantiacus. Spectrochim Acta A Mol Biol Spectrosc 56:2001–2010

  23. Miyatake T, Tamiaki H (2005) Self-aggregates of bacteriochlorophylls-c, d and e in a light-harvesting antenna system of green photosynthetic bacteria: effect of stereochemistry at the chiral 3-(1-hydroxyethyl) group on the supramolecular arrangement of chlorophyllous pigments. J Photoch Photobio C 6:89–107

  24. Miyatake T, Tamiaki H (2010) Self-aggregates of natural chlorophylls and their synthetic analogs in aqueous media for making light-harvesting systems. Coord Chem Rev 254:2593–2602

  25. Montano GA, Wu HM, Lin S, Brune DC, Blankenship RE (2003) Isolation and characterization of the B798 light-harvesting baseplate from the chlorosomes of Chloroflexus aurantiacus. Biochemistry 42:10246–10251

  26. Oostergetel GT, Reus M, Gomez Maqueo Chew A, Bryant DA, Boekema EJ, Holzwarth AR (2007) Long-range organization of bacteriochlorophyll in chlorosomes of Chlorobium tepidum investigated by cryo-electron microscopy. FEBS Lett 581:5435–5439

  27. Oostergetel GT, van Amerongen H, Boekema EJ (2010) The chlorosome: a prototype for efficient light-harvesting in photosynthesis. Photosynth Res 104:245–255

  28. 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

  29. Pedersen MO, Underhaug J, Dittmer J, Miller M, Nielsen NC (2008) The three-dimensional structure of CsmA: a small antenna protein from the green sulfur bacterium Chlorobium tepidum. FEBS Lett 582:2869–2874

  30. Polivka T, Frank HA (2010) Molecular factors controlling photosynthetic light-harvesting by carotenoids. Acc Chem Res 43:1125–1134

  31. Psencik J, Ma YZ, Arellano JB, Garcia-Gil J, Holzwarth AR, Gillbro T (2002) Excitation energy transfer in chlorosomes of Chlorobium phaeobacteroides strain CL1401: the role of carotenoids. Photosynth Res 71:5–18

  32. Psencik J, Ma YZ, Arellano JB, Hala J, Gillbro T (2003) Excitation energy transfer dynamics and excited-state structure in chlorosomes of Chlorobium phaeobacteroides. Biophys J 84:1161–1179

  33. Psencik J, Ikonen TP, Laurinmäki P, Merckel MC, Butcher SJ, Serimaa RE, Tuma R (2004) Lamellar organization of pigments in chlorosomes, the light-harvesting complexes of green photosynthetic bacteria. Biophys J 87:1165–1172

  34. Psencik J, Arellano JB, Ikonen TP, Borrego CM, Laurinmäki PA, Butcher SJ, Serimaa RE, Tuma R (2006) Internal structure of chlorosomes from brown-colored Chlorobium species and the role of carotenoids in their assembly. Biophys J 91:1433–1440

  35. Psencik J, Collins AM, Liljeroos L, Torkkeli M, Laurinmäki P, Ansink HM, Ikonen TP, Serimaa RE, Blankenship RE, Tuma R, Butcher SJ (2009) Structure of chlorosomes from the green filamentous bacterium Chloroflexus aurantiacus. J Bacteriol 191:6701–6708

  36. Sakuragi Y, Frigaard NU, Shimada K, Matsuura K (1999) Association of bacteriochlorophyll a with the CsmA protein in chlorosomes of the photosynthetic green filamentous bacterium Chloroflexus aurantiacus. BBA-Bioenergetics 1413:172–180

  37. Sorensen PG, Cox RP, Miller M (2008) Chlorosome lipids from Chlorobium tepidum: characterization and quantification of polar lipids and wax esters. Photosynth Res 95:191–196

  38. Stanier RY, Smith JHC (1960) The chlorophylls of green bacteria. Biochim Biophys Acta 41:478–484

  39. Steensgaard DB, Wackerbarth H, Hildebrandt P, Holzwarth AR (2000) Diastereoselective control of bacteriochlorophyll e aggregation. 3(1)-S-BChl e is essential for the formation of chlorosome-like aggregates. J Phys Chem B 104:10379–10386

  40. Takaichi S, Wang ZY, Umetsu M, Nozawa T, Shimada K, Madigan MT (1997) New carotenoids from the thermophilic green sulfur bacterium Chlorobium tepidum: 1′,2′-dihydro-gamma-carotene, 1′,2′-dihydrochlorobactene, and OH-chlorobactene glucoside ester, and the carotenoid composition of different strains. Arch Microbiol 168:270–276

  41. Uehara K, Mimuro M, Ozaki Y, Olson JM (1994) The formation and characterization of the in vitro polymeric aggregates of bacteriochlorophyll c homologs from Chlorobium limicola in aqueous suspension in the presence of monogalactosyl diglyceride. Photosynth Res 41:235–243

  42. Umetsu M, Seki R, Kadota T, Wang ZY, Adschiri T, Nozawa T (2003) Dynamic exchange properties of the antiparallel bacteriochlorophyll c dimers. J Phys Chem B 107:9876–9882

  43. van Stokkum IHM, Larsen DS, van Grondelle R (2004) Global and target analysis of time-resolved spectra. BBA-Bioenergetics 1657:82–104

  44. Weber S (1988) Determination of stabilised, added astaxanthin in fish feeds and premixes with HPLC. In: Keller HE (ed) Analytical methods for vitamins and carotenoids in feeds, pp 59–61. Roche Publication No. 2264, Basel

  45. Zupcanova A, Arellano JB, Bina D, Kopecky J, Psencik J, Vacha F (2008) The length of esterifying alcohol affects the aggregation properties of chlorosomal bacteriochlorophylls. Photochem Photobiol 84:1187–1194

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Acknowledgments

This study was supported by the Czech Ministry of Education, Youth and Sports (projects MSM0021620835, MSM6007665808, AV0Z50510513), Czech Science Foundation (projects 206/09/0375, 202/09/H041, 202/09/1330), and Spanish Ministry of Science and Innovation (AVCR-CSIC joint project 2008CZ0004). The authors would like to thank Ivana Hunalova, Frantisek Matousek, and Anita Zupcanova for their help with pigment isolation.

Author information

Correspondence to J. Pšenčík.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Figure S1: Correction of the absorption spectra of BChl c-astaxanthin assemblies for light-scattering. Spectra were measured at a position further (solid line) and closer (dotted line) to the detector of the spectrophotometer. The difference between them were rescaled to fit the uncorrected spectra above 850 nm which reflected the scattering part of the spectra (dashed line). The corrected spectrum (dash-dot line) was obtained by subtracting the scattering part from the solid line. (PDF 21 kb)

Figure S2: Transient absorption spectra of BChl c assemblies with astaxanthin (molar ratio 0.35:1 astaxanthin-to-BChl c, blue lines) and BChl c dimers with lecithin (green lines). All spectra were measured at 2.1 ps after selective excitation of BChl c at 710 nm. The inset shows an enlargement of the 450-600 nm region. The inverted absorption spectrum of astaxanthin in the assemblies, reproduced from Fig. 3 and scaled to fit, is shown for the sake of comparison (black line, inset). The difference spectrum (red line) reveals a shift of the Qy band of BChl c and a small negative signal in the 450-600 nm region. This small signal can be caused entirely by a change in the BChl–BChl excitonic interaction between the two samples, or by an additional excitonic interaction between BChl c and astaxanthin resulting in bleaching of the astaxanthin upon BChl c excitation. The spectra were smoothed by a 10 point (± 2.5 nm) adjacent averaging. (PDF 28 kb)

Appendix: Transient data analysis

Appendix: Transient data analysis

We assume simple kinetic model for excited state population N i

$$ \dot{N}_{i} (t) = \sum\limits_{i} {\left( { - k_{ij} N_{i} (t) + k_{ji} N_{j} (t)} \right)} $$
(1)

where k ij is the rate constant for energy/population transfer from one state i to the another state j and dot represents derivative with respect to time. By solving set of Eq. 1 (one for each state), and considering that each state contributes to transient signal proportionally to its population, we get the following model for describing transient absorption data

$$ \Updelta A(t) = \sum\limits_{j} {\left( {\sum\limits_{i} {K_{ji} \varepsilon_{i} N_{i} } (0)} \right)\exp ( - k_{j} t)} $$
(2)

where indices i and j iterates over all energy states present in the sample, k j is the overall rate constant of the state j (\( k_{j} = \sum\limits_{i} {k_{ij} } \)), ε is a spectral profile associated with the given state, and coefficients K ij describe the energy transfer between states, typically \( K_{ij} = \frac{{k_{ij} }}{{k_{j} - k_{i} }} \) or product of similar terms. We can then perform global fitting analysis to find the overall rate constants and corresponding pre-exponential factors (i.e. the DAS)

$$ \Updelta A(t) = \sum\limits_{j} {DAS_{j} \exp ( - k_{j} t)} $$
(3)

Let’s assume in accordance with the multi-exponential model that relaxation pathways from an excited state are the same regardless of its population. This is of course an approximation, because relaxation within the BChl c manifold is likely to be influenced by singlet–singlet annihilation, which the model does not describe. It follows that the transient signal consists of parts proportional to the initial population of states directly excited by pump pulse (the other way of summation in Eq. 2)

$$ \Updelta A(t) = \sum\limits_{i} {\left( {\varepsilon_{i} \sum\limits_{j} {K_{ji} \exp ( - k_{j} t)} } \right)N_{i} (0)} = \sum\limits_{i} {\Updelta a_{i} N_{i} (0)} $$
(4)

If we perform at least P measurements with different and known ratio of the initial population of the excited states, where P is the number of the initially excited states (with non-zero N i (0)), then we can determine the parts of the transient data corresponding to the relaxation from each of the initially excited states (Δa i in Eq. 4). Note that these parts should be the same in all measurements, except that their weights N i (0) differ. Here we assume that only the S2 state of astaxanthin and Soret band of BChl c are directly excited, and that the transient data separates into two parts: relaxation following excitation of astaxanthin, and BChl c, respectively. Therefore, we need to perform two transient absorption measurements with a different ratio between initial populations of the S2 state of astaxanthin and the Soret band of BChl c. One theoretically possible way to change the excited state population is to use one excitation wavelength and two samples with different astaxanthin-to-BChl c molar ratios. However, different sample composition in this case results in a different way of assembly, and therefore, in different absorption spectra, breaking the condition that relaxation from excited state is the same. The other way is to use one sample and two different excitation wavelengths (close enough to excite the same electronic transitions and far enough to achieve significant change of excited populations ratio). In this case, we populate different parts from the inhomogeneously broadened electronic transition peaks; however, this should not influence the relaxation from the higher excited state significantly. We measured the transient data after excitation at two different wavelengths, 450 and 490 nm. Therefore, we can rewrite Eq. 4 describing the global decay at two excitation wavelength as

$$ \begin{gathered} \Updelta A_{450} = \Updelta a_{\text{BChl}} N_{450}^{\text{BChl}} (0) + \Updelta a_{\text{Car}} N_{450}^{\text{Car}} (0) \hfill \\ \Updelta A_{490} = \Updelta a_{\text{BChl}} N_{490}^{\text{BChl}} (0) + \Updelta a_{\text{Car}} N_{490}^{\text{Car}} (0) \hfill \\ \end{gathered} $$
(5)

where ΔA 450 and ΔA 490 represent transient data after excitation at 450 and 490 nm, respectively.

The initial excited state populations N λ molecule (0) could be determined directly from steady-state absorption spectra corrected for scattering, and energy in the pump pulse. From the two equations, two unknown matrices Δa BChl and Δa Car, which represent BChl c and astaxanthin part of the transient data, could be calculated (Fig. 6).

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Alster, J., Polívka, T., Arellano, J.B. et al. Self-assembly and energy transfer in artificial light-harvesting complexes of bacteriochlorophyll c with astaxanthin. Photosynth Res 111, 193–204 (2012). https://doi.org/10.1007/s11120-011-9670-0

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Keywords

  • Light-harvesting
  • Chlorosomes
  • Self-assembly
  • Bacteriochlorophyll aggregates
  • Astaxanthin