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Transient EPR: using spin polarization in sequential radical pairs to study electron transfer in photosynthesis

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Abstract

The light induced electron transfer in photosynthesis generates a series of sequential spin polarized radical pairs, and transient electron paramagnetic resonance (TREPR) is ideally suited to study the lifetimes and physical and electronic structures of these radical pairs. In this article, the basic principles of TREPR are outlined with emphasis on the electron spin polarization (ESP) that develops during the electron transfer process. Examples of the analysis of TREPR data are given to illustrate the information that can be obtained. Recent applications of the technique to study the functionality of reaction centers, light-induced structural changes, and protein–cofactor interactions are reviewed.

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References

  • Agalarov R, Byrdin M, Rappaport F, Shen GZ, Bryant DA, van der Est A, Golbeck J (2008) Removal of the PsaF polypeptide biases electron transfer in favor of the PsaB branch of cofactors in Triton X-100 photosystem I complexes from Synechococcus sp. PCC 7002. Photochem Photobiol 84:1371–1380. doi:10.1111/j.1751-1097.2008.00457.x

    Article  CAS  PubMed  Google Scholar 

  • Bittl R, Weber S (2005) Transient radical pairs studied by time-resolved EPR Biochim. Biophys. Acta-Bioenerg 1707:117–126. doi:10.1016/j.bbabio.2004.03.012

    Article  CAS  Google Scholar 

  • Bock CH, van der Est AJ, Brettel K, Stehlik D (1989) Nanosecond electron transfer kinetics in photosystem I as obtained from transient EPR at room temperature. FEBS Lett 247:91–96. doi:10.1016/0014-5793(89)81247-5

    Article  CAS  Google Scholar 

  • Boudreaux B, MacMillan F, Teutloff C, Agalarov R, Gu FF, Grimaldi S, Bittl R, Brettel K, Redding K (2001) Mutations in both sides of the photosystem I reaction center identify the phylloquinone observed by electron paramagnetic resonance spectroscopy. J Biol Chem 276:37299–37306. doi:10.1074/jbc.M102327200

    Article  CAS  PubMed  Google Scholar 

  • Brettel K (1988) Electron transfer from A1 − to an iron-sulfur center with t1/2 = 200 ns at room temperature in photosystem I characterization by flash absorption spectroscopy. FEBS Lett 239:93–98. doi:10.1016/0014-5793(88)80552-0

    Article  CAS  Google Scholar 

  • Dubinski AA, Perekhodtsev GD, Poluektov OG, Rajh T, Thurnauer MC (2002) Analytical treatment of EPR spectra of weakly coupled spin-correlated radical pairs in disordered solids: application to the charge-separated state in TiO2 nanoparticles. J Phys Chem B 106:938–944. doi:10.1021/jp013276h

    Article  CAS  Google Scholar 

  • Guergova-Kuras M, Boudreaux B, Joliot A, Joliot P, Redding K (2001) Evidence for two active branches for electron transfer in photosystem I. Proc Natl Acad Sci USA 98:4437–4442. doi:10.1073/pnas.081078898

    Article  CAS  PubMed  Google Scholar 

  • Heinent U, Utschig LM, Poluektov OG, Link G, Ohmes E, Kothe G (2007) Structure of the charge separated state P(865)(+)Q(A)(−) in the photosynthetic reaction centers of Rhodobacter sphaeroides by quantum beat oscillations and high-field electron paramagnetic resonance: evidence for light-induced Q(A)(−) reorientation. J Am Chem Soc 129:15935–15946. doi:10.1021/ja075065h

    Article  CAS  Google Scholar 

  • Hoff AJ (1984) Electron-spin polarization of photosynthetic reactants Q. Rev Biophys 17:153–282

    Article  CAS  Google Scholar 

  • Hore PJ (1996) Transfer of spin correlation between radical pairs in the initial steps of photosynthetic energy conversion. Mol Phys 89:1195–1202. doi:10.1080/002689796173598

    CAS  Google Scholar 

  • Hulsebosch RJ, Borovykh IV, Paschenko SV, Gast P, Hoff AJ (1999) Radical pair dynamics and interactions in quinone-reconstituted photosynthetic reaction centers of Rb. sphaeroides R26: a multifrequency magnetic resonance study. J Phys Chem B 103:6815–6823. doi:10.1021/jp990677c

    Article  CAS  Google Scholar 

  • Ishikita H, Knapp EW (2003) Redox potential of quinones in both electron transfer branches of photosystem. J Biol Chem 278:52002–52011. doi:10.1074/jbc.M306434200

    Article  CAS  PubMed  Google Scholar 

  • Jordan P, Fromme P, Witt HT, Klukas O, Saenger W, Krauss N (2001) Three-dimensional structure of cyanobacterial photosystem I at 25 angstrom resolution. Nature 411:909–917. doi:10.1038/35082000

    Article  CAS  PubMed  Google Scholar 

  • Kandrashkin Y, van der Est A (2001) A new approach to determining the geometry of weakly coupled radical pairs from their electron spin polarization patterns. Spectrochim Acta A 57:1697–1709. doi:10.1016/S1386-1425(01)00436-X

    Article  CAS  Google Scholar 

  • Kandrashkin YE, van der Est A (2007) Time-resolved EPR spectroscopy of photosynthetic reaction centers: from theory to experiment. Appl Magnes Res 31:105–122

    Article  CAS  Google Scholar 

  • Kandrashkin YE, Salikhov KM, Stehlik D (1997) Spin dynamics and EPR spectra of consecutive spin-correlated radical pairs. Model calculations. Appl Magn Res 12:141–166

    Article  CAS  Google Scholar 

  • Kandrashkin YE, Salikhov KM, van der Est A, Stehlik D (1998) Electron spin polarization in consecutive spin-correlated radical pairs: application to short-lived and long-lived precursors in type 1 photosynthetic reaction centres. Appl Magnes Res 15:417–447

    Article  CAS  Google Scholar 

  • Karyagina I, Golbeck JH, Srinivasan N, Stehlik D, Zimmermann H (2006) Single-sided hydrogen bonding to the quinone cofactor in photosystem I probed by selective C-13-labelled naphthoquinones and transient EPR. Appl Magnes Res 30:287–310

    Article  CAS  Google Scholar 

  • Karyagina I, Pushkar Y, Stehlik D, van der Est A, Ishikita H, Knapp EW, Jagannathan B, Agalarov R, Golbeck JH (2007) Contributions of the protein environment to the midpoint potentials of the A(1) phylloquinones and the F-x iron-sulfur cluster in photosystem I. Biochemistry 46:10804–10816

    Article  CAS  PubMed  Google Scholar 

  • Kim SS, Weissman SI (1976) Detection of transient electron-paramagnetic resonance. J Magn Res 24:167–169

    Google Scholar 

  • Kleinfeld D, Okamura MY, Feher G (1984) Electron-transfer kinetics in photosynthetic reaction centers cooled to cryogenic temperatures in the charge-separated state—evidence for light-induced structural-changes. Biochemistry 23:5780–5786

    Article  CAS  PubMed  Google Scholar 

  • Klette R, Torring JT, Plato M, Möbius K, Bönigk B, Lubitz W (1993) Determination of the G tensor of the primary donor cation radical in single-crystals of Rhodobacter sphaeroides R-26 reaction centers by 3-Mm high-field EPR. J Phys Chem 97:2015–2020. doi:10.1021/j100111a047

    Article  CAS  Google Scholar 

  • Kothe G, Norris JR, Poluektov OG, Thurnauer MC (2008) High-time resolution electron paramagnetic resonance study of quantum beat oscillations observed photosynthetic reaction center proteins. In: Aartsma T, Matysik J (eds) Biophysical techniques in photosynthesis, vol. II. Advances in photosynthesis and respiration, vol. 26. Springer, Dordrecht, The Netherlands, pp 305–323

  • Levanon H (1996) Time-resolved electron paramagnetic resonance spectroscopy—principles and applications. In: Amesz J, Hoff AJ (eds) Biophysical techniques in photosynthesis. Advances in photosynthesis and respiration, vol 3. Kluwer Academic Publishers, Dordrecht, The Netherlands, pp 211–233

  • Li YJ, van der Est A, Lucas MG, Ramesh VM, Gu FF, Petrenko A, Lin S, Webber AN, Rappaport F, Redding K (2006) Directing electron transfer within photosystem I by breaking H-bonds in the cofactor branches. Proc Natl Acad Sci USA 103:2144–2149. doi:10.1073/pnas.0506537103

    Article  CAS  PubMed  Google Scholar 

  • Lubitz W, Lendzian F, Bittl R (2002) Radicals, radical pairs and triplet states in photosynthesis. Acc Chem Res 35:313–320. doi:10.1021/ar000084g

    Article  CAS  PubMed  Google Scholar 

  • Miyamoto R, Nlino H, Kondo T, Itoh S, Oh-Oka H (2008) An electron spin-polarized signal of the P800(+)A(1)(Q)(−) state in the homodimeric reaction center core complex of Heliobacterium modesticaldum. Biochemistry 47:4386–4393. doi:10.1021/bi701612v

    Article  CAS  PubMed  Google Scholar 

  • Möbius K (2000) Primary processes in photosynthesis: what do we learn from high-field EPR spectroscopy? Chem Soc Rev 29:129–139. doi:10.1039/a706426h

    Article  Google Scholar 

  • Morris AL, Snyder SW, Zhang YN, Tang J, Thurnauer MC, Dutton PL, Robertson DE, Gunner MR (1995) Electron-spin polarization model applied to sequential electron-transfer in iron-containing photosynthetic bacterial reaction centers with different quinones as Q(a). J Phys Chem 99:3854–3866. doi:10.1021/j100011a063

    Article  CAS  Google Scholar 

  • Norris JR, Morris AL, Thurnauer MC, Tang J (1990) A general-model of electron-spin polarization arising from the interactions within radical pairs. J Chem Phys 92:4239–4249. doi:10.1063/1.457782

    Article  CAS  Google Scholar 

  • Pedersen JB (1979) Determination of the primary reactions of photosynthesis transient ESR signals. FEBS Lett 97:305–310

    Article  CAS  Google Scholar 

  • Poluektov OG, Paschenko SV, Utschig LM, Lakshmi KV, Thurnauer MC (2005) Bidirectional electron transfer in photosystem I: direct evidence from high-frequency time-resolved EPR spectroscopy. J Am Chem Soc 127:11910–11911. doi:10.1021/ja053315t

    Article  CAS  PubMed  Google Scholar 

  • Prisner TF, van der Est A, Bittl R, Lubitz W, Stehlik D, Möbius K (1995) Time-resolved W-Band (95 GHz) EPR spectroscopy of Zn-substituted reaction centers of Rhodobacter sphaeroides R–26. Chem Phys 194:361–370. doi:10.1016/0301-0104(95)00016-H

    Article  CAS  Google Scholar 

  • Prisner T, Rohrer M, MacMillan F (2001) Pulsed EPR spectroscopy: biological applications. Annu Rev Phys Chem 52:279–313. doi:10.1146/annurev.physchem.52.1.279

    Article  CAS  PubMed  Google Scholar 

  • Purton S, Stevens DR, Muhiuddin IP, Evans MCW, Carter S, Rigby SEJ, Heathcote P (2001) Site-directed mutagenesis of PsaA residue W693 affects phylloquinone binding and function in the photosystem I reaction center of Chlamydomonas reinhardtii. Biochemistry 40:2167–2175. doi:10.1021/bi0019489

    Article  CAS  PubMed  Google Scholar 

  • Pushkar YN, Golbeck JH, Stehlik D, Zimmermann H (2004) Asymmetric hydrogen-bonding of the quinone cofactor in photosystem I probed by C-13-labeled naphthoquinones. J Phys Chem B 108:9439–9448. doi:10.1021/jp0361879

    Article  CAS  Google Scholar 

  • Pushkar YN, Ayzatulin O, Stehlik D (2005) Transient and pulsed EPR study of O-17-substituted methyl-naphthoquinone as radical anion in the A(1) binding site of photosystem I and in frozen solution. Appl Magn Res 28:195–211

    Article  CAS  Google Scholar 

  • Redding K, van der Est A (2006) The directionality of electron transport in photosystem I. In: Golbeck J (ed) Photosystem I. The light-driven plastocyanin: ferredoxin oxidoreductase. Advances in photosynthesis and respiration, vol. 24. Springer, Dordrecht, The Netherlands, pp 413–437

  • Savitsky A, Möbius K (2006) Photochemical reactions and photoinduced electron-transfer processes in liquids, frozen solutions, and proteins as studied by multifrequency time-resolved EPR spectroscopy. Helv Chim Acta 89:2544–2589. doi:10.1002/hlca.200690232

    Article  CAS  Google Scholar 

  • Savitsky A, Dubinskii AA, Flores M, Lubitz W, Möbius K (2007) Orientation-resolving pulsed electron dipolar high-field EPR spectroscopy on disordered solids: I Structure of spin-correlated radical pairs in bacterial photosynthetic reaction centers. J Phys Chem B 111:6245–6262. doi:10.1021/jp070016c

    Article  CAS  PubMed  Google Scholar 

  • Schins JM, Prins P, Grozema FC, Abellon RD, de Haas MP, Siebbeles LDA (2005) Development of a microwave transmission setup for time-resolved measurements of the transient complex conductivity in bulk samples. Rev Sci Instrum 76:084703. doi:10.1063/1.2008978

    Article  CAS  Google Scholar 

  • Semenov AY, Vassiliev IR, van der Est A, Mamedov MD, Zybailov B, Shen GZ, Stehlik D, Diner BA, Chitnis PR, Golbeck JH (2000) Recruitment of a foreign quinone into the A(1) site of photosystem I—altered kinetics of electron transfer in phylloquinone biosynthetic pathway mutants studied by time-resolved optical, EPR, and electrometric techniques. J Biol Chem 275:23429–23438. doi:10.1074/jbc.M000508200

    Article  CAS  PubMed  Google Scholar 

  • Setif P, Brettel K (1993) Forward electron transfer from phylloquinone A1 to iron-sulfur centers in spinach photosystem I. Biochemistry 32:7846–7854

    Article  CAS  PubMed  Google Scholar 

  • Shen GZ, Antonkine ML, van der Est A, Vassiliev IR, Brettel K, Bittl R, Zech SG, Zhao JD, Stehlik D, Bryant DA, Golbeck JH (2002) Assembly of photosystem I. II Rubredoxin is required for the in vivo assembly of Fx in Synechococcus sp. PCC 7002 as shown by optical and EPR spectroscopy. J Biol Chem 277:20355–20366. doi:10.1074/jbc.M201104200

    Article  CAS  PubMed  Google Scholar 

  • Snyder SW, Morris AL, Bondeson SR, Norris JR, Thurnauer MC (1993) Electron-spin polarization in sequential electron-transfer—an example from iron-containing photosynthetic bacterial reaction center proteins. J Am Chem Soc 115:3774–3775. doi:10.1021/ja00062a053

    Article  CAS  Google Scholar 

  • Stehlik D (2006) Transient EPR spectroscopy as applied to light-induced functional intermediates along the electron transfer pathway in photosystem I. In: Golbeck J (ed) Photosystem I. The light-driven plastocyanin: ferredoxin oxidoreductase. Advances in photosynthesis and respiration, vol 24. Springer, Dordrecht, The Netherlands, pp 361–386

  • Stehlik D, Möbius K (1997) New EPR methods for investigating photoprocesses with paramagnetic intermediates. Annu Rev Phys Chem 48:745–784. doi:10.1146/annurev.physchem.48.1.745

    Article  CAS  PubMed  Google Scholar 

  • Stehlik D, Bock CH, Thurnauer MC (1989) Transient EPR-spectroscopy of photoinduced electronic spin states in rigid matrices. In: Hoff AJ (ed) Advanced EPR in biology and biochemistry. Elsevier, Amsterdam, pp 371–403

    Google Scholar 

  • Stowell MHB, McPhillips TM, Rees DC, Soltis SM, Abresch E, Feher G (1997) Light-induced structural changes in photosynthetic reaction center: implications for mechanism of electron-proton transfer. Science 276:812–816. doi:10.1126/science.276.5313.812

    Article  CAS  PubMed  Google Scholar 

  • Synder SW, Thurnauer MC (1993) The photosynthetic reaction center. In: Deisenhofer J, Norris JR (eds) Electron spin polarization in photosynthetic reaction centers, vol II. Academic Press, San Diego, pp 285–329

    Google Scholar 

  • Tang J, Bondeson S, Thurnauer MC (1996) Effects of sequential electron transfer on electron spin polarized transient EPR spectra at high fields. Chem Phys Lett 253:293–298. doi:10.1016/0009-2614(96)00176-5

    Article  CAS  Google Scholar 

  • Tang J, Utschig LM, Poluektov O, Thurnauer MC (1999) Transient W-band EPR study of sequential electron transfer in photosynthetic bacterial reaction centers. J Phys Chem B 103:5145–5150. doi:10.1021/jp990236t

    Article  CAS  Google Scholar 

  • Thurnauer MC, Norris JR (1980) An electron-spin echo phase-shift observed in photosynthetic algae—possible evidence for dynamic radical pair interactions. Chem Phys Lett 76:557–561

    Article  CAS  Google Scholar 

  • Utschig LM, Greenfield SR, Tang J, Laible PD, Thurnauer MC (1997) Influence of iron-removal procedures on sequential electron transfer in photosynthetic bacterial reaction centers studied by transient EPR spectroscopy. Biochemistry 36:8548–8558. doi:10.1021/bi9630319

    Article  CAS  PubMed  Google Scholar 

  • van der Est A (2001) Light-induced spin polarization in type I photosynthetic reaction centres. Biochim Biophys Acta Bioenerg 1507:212–225. doi:10.1016/S0005-2728(01)00204-3

    Article  Google Scholar 

  • van der Est A, Bittl R, Abresch EC, Lubitz W, Stehlik D (1993) Transient EPR spectroscopy of perdeuterated Zn-substituted reaction centers of Rhodobacter sphaeroides R-26. Chem Phys Lett 212:561–568. doi:10.1016/0009-2614(93)85487-9

    Article  Google Scholar 

  • van der Est A, Bock C, Golbeck J, Brettel K, Sétif P, Stehlik D (1994) Electron transfer from acceptor A1 to the iron-sulfur centers in photosystem I as studied by transient EPR spectroscopy. Biochemistry 33:11789–11797. doi:10.1021/bi00205a015

    Article  PubMed  Google Scholar 

  • van der Est A, Hager-Braun C, Leibl W, Hauska G, Stehlik D (1998) Transient electron paramagnetic resonance spectroscopy on green-sulfur bacteria and heliobacteria at two microwave frequencies. Biochim Biophys Acta 1409:87–98. doi:10.1016/S0005-2728(98)00152-2

    Article  PubMed  Google Scholar 

  • van der Est A, Valieva AI, Kandrashkin YE, Shen GZ, Bryant DA, Golbeck JH (2004) Removal of PsaF alters forward electron transfer in photosystem I: evidence for fast reoxidation of Q(K)-A in subunit deletion mutants of Synechococcus sp. PCC 7002. Biochemistry 43:1264–1275

    Article  CAS  Google Scholar 

  • Xu W, Chitnis P, Valieva A, van der Est A, Pushkar YN, Krzystyniak M, Teutloff C, Zech SG, Bittl R, Stehlik D, Zybailov B, Shen GZ, Golbeck JH (2003a) Electron transfer in cyanobacterial photosystem I–II. Physiological and spectroscopic characterization of site-directed mutants in a putative electron transfer pathway from A0 through A1 to Fx. J Biol Chem 278:27864–27875. doi:10.1074/jbc.M302962200

    Article  CAS  PubMed  Google Scholar 

  • Xu W, Chitnis PR, Valieva A, van der Est A, Brettel K, Guergova-Kuras M, Pushkar YN, Zech SG, Stehlik D, Shen GZ, Zybailov B, Golbeck JH (2003b) Electron transfer in cyanobacterial photosystem I–II. Determination of forward electron transfer rates of site-directed mutants in a putative electron transfer pathway from A(0) through A(1) to F-x. J Biol Chem 278:27876–27887. doi:10.1074/jbc.M302965200

    Article  CAS  PubMed  Google Scholar 

  • Zybailov B, van der Est A, Zech SG, Teutloff C, Johnson TW, Shen GZ, Bittl R, Stehlik D, Chitnis PR, Golbeck JH (2000) Recruitment of a foreign quinone into the A1 site of photosystem I–II. Structural and functional characterization of phylloquinone biosynthetic pathway mutants by electron paramagnetic resonance and electron-nuclear double resonance spectroscopy. J Biol Chem 275:8531–8539. doi:10.1074/jbc.275.12.8531

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This article is dedicated to the memory of Dietmar Stehlik who introduced me to both TREPR and Photosynthesis. Financial support was provided by the Natural Sciences and Engineering Research Council, Canada.

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  1. Department of Chemistry, Brock University, Saint Catharines, ON, Canada

    Art van der Est

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van der Est, A. Transient EPR: using spin polarization in sequential radical pairs to study electron transfer in photosynthesis. Photosynth Res 102, 335–347 (2009). https://doi.org/10.1007/s11120-009-9411-9

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