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Correlation of the cytochrome c 2 content of cyanobacterial Photosystem II with the EPR properties of the oxygen-evolving complex

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Abstract

The Mn4 cluster of PS II advances through a series of oxidation states (S states) that catalyze the breakdown of water to dioxygen in the oxygen-evolving complex. The present study describes the engineering and purification of highly active PS II complexes from mesophilic His-tagged Synechocystis PCC 6803 and purification of PS II core complexes from thermophilic wild-type Synechococcus lividus with high levels of the extrinsic polypeptide, cytochrome c 550. The g = 4.1 S2 state EPR signal, previously not characterized in untreated cyanobacterial PS II, is detected in high yields in these PS II preparations. We present a complete characterization of the g = 4.1 state in cyanobacterial His-tagged Synechocystis PCC 6803 PS II and S. lividus PS II. Also presented are a determination of the stoichiometry of cytochrome c 550 bound to His-tagged Synechocystis PCC 6803 PS II and analytical ultracentrifugation results which indicate that cytochrome c 550 is a monomer in solution. The temperature-dependent multiline to g = 4.1 EPR signal conversion observed for the S2 state in cyanobacterial PS II with high cytochrome c 550 content is very similar to that previously found for spinach PS II. In spinach PS II, the formation of the S2 state g = 4.1 EPR signal has been found to correlate with the binding of the extrinsic 17 and 23 kDa polypeptides. The finding of a similar correlation in cyanobacterial PS II with the binding of cytochrome c 550 suggests a functional homology between cytochrome c 550 and the 17 and 23 kDa extrinsic proteins of spinach PS II.

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References

  • Åhrling KA, Peterson S and Styring S (1997) An oscillating manganese electron paramagnetic resonance signal from the S-0 state of the oxygen evolving complex in Photosystem II. Biochemistry 36: 13148-13152

    Article  PubMed  CAS  Google Scholar 

  • Beck WF and Brudvig GW (1988) Ligand-substitution reactions of the O2-evolving center of Photosystem II. Chem Scr 28A: 93-98

    Google Scholar 

  • Boussac A and Rutherford AW (2000) Comparative study of the g = 4.1 EPR signals in the S-2 state of Photosystem II. Biochim Biophys Acta 1457: 145-156

    Article  PubMed  CAS  Google Scholar 

  • Boussac A, Girerd JJ and Rutherford AW (1996) Conversion of the spin state of the manganese complex in Photosystem II induced by near-infrared light. Biochemistry 35: 6984-6989

    Article  PubMed  CAS  Google Scholar 

  • Boussac A, Kuhl H, Un S, Rogner M and Rutherford AW (1998a) Effect of near-infrared light on the S-2 state of the manganese complex of Photosystem II from Synechococcus elongatus. Biochemistry 37: 8995-9000

    Article  PubMed  CAS  Google Scholar 

  • Boussac A, Un S and Horner O (1998b) High-spin states (S = 5/2) of the Photosystem II manganese complex. Biochemistry 37: 4001-4007

    Article  PubMed  CAS  Google Scholar 

  • Boussac A, Sugiura M, Inoue Y and Rutherford AW (2000) EPR study of the oxygen evolving complex in His-tagged Photosystem II from the cyanobacterium Synechococcus elongatus. Biochemistry 39: 13788-13799

    Article  PubMed  CAS  Google Scholar 

  • Bricker TM (1990) The structure and function of CPa-1 and CPa-2 in Photosystem II. Photosynth Res 24: 1-13

    Article  CAS  Google Scholar 

  • Bricker TM, Morvant J, Masri N, Sutton HM and Frankel LK (1998) Isolation of a highly active Photosystem II preparation from Synechocystis 6803 using a histidine-tagged mutant of CP47. Biochim Biophys Acta 1409: 50-57

    Article  PubMed  CAS  Google Scholar 

  • Brudvig GW, Casey JL and Sauer K (1983) The effect of temperature on the formation and decay of the multiline electronparamagnetic-resonance signal species associated with photosynthetic oxygen evolution. Biochim Biophys Acta 723: 366-371

    Article  CAS  Google Scholar 

  • Campbell KA, Gregor W, Pham DP, Peloquin JM, Debus RJ and Britt RD (1998a) The 23 and 17 kDa extrinsic proteins of Photosystem II modulate the magnetic properties of the S1-state manganese cluster. Biochemistry 37: 5039-5045

    Article  PubMed  CAS  Google Scholar 

  • Campbell KA, Peloquin JM, Pham DP, Debus RJ and Britt RD (1998b) Parallel polarization EPR detection of an S-1-state 'multiline' EPR signal in Photosystem II particles from Synechocystis sp. PCC 6803. J Am Chem Soc 120: 447-448

    Article  CAS  Google Scholar 

  • Casey JL and Sauer K (1984) Electron-paramagnetic-resonance detection of a cryogenically photogenerated intermediate in photosynthetic oxygen evolution. Biochim Biophys Acta 767: 21-28

    Article  CAS  Google Scholar 

  • Chua N-H (1980) Electrophoretic analysis of chloroplast proteins. Meth Enzymol 69: 434-446

    CAS  Google Scholar 

  • Cramer WA, Theg SM and Widger WR (1986) On the structure and function of cytochrome b-559. Photosynth Res 10: 393-403

    Article  CAS  Google Scholar 

  • de Paula JC, Beck WF, Miller A-F, Wilson RB and Brudvig GW (1987) Studies of the manganese site of Photosystem II by electron paramagnetic resonance spectroscopy. J Chem Soc Faraday Trans 83: 3635-3651

    Article  Google Scholar 

  • de Paula JC, Innes JB and Brudvig GW (1985) Electron-transfer in Photosystem II at cryogenic temperatures. Biochemistry 24: 8114-8120

    Article  PubMed  CAS  Google Scholar 

  • de Paula JC, Li PM, Miller A-F, Wu BW and Brudvig GW (1986) Effect of the 17-kDa and 23-kDa polypeptides, calcium and chloride on electron-transfer in Photosystem II. Biochemistry 25: 6487-6494

    Article  PubMed  CAS  Google Scholar 

  • Debus RJ (1992) Themanganese and calcium ions of photosynthetic oxygen evolution. Biochim Biophys Acta 1102: 269-352

    Article  PubMed  CAS  Google Scholar 

  • Dismukes GC and Siderer Y (1981) Intermediates of a polynuclear manganese center involved in photosynthetic oxidation of water. Proc Natl Acad Sci USA 78: 274-278

    Article  PubMed  CAS  Google Scholar 

  • Frazao C, Enguita FJ, Coelho R, Sheldrick GM, Navarro JA, Hervas M, De la Rosa MA and Carrondo MA (2001) Crystal structure of low-potential cytochrome c549 from Synechocystis sp. PCC 6803 at 1.21 Å resolution. J Biol Inorg Chem 6: 324-332

    Article  PubMed  CAS  Google Scholar 

  • Haddy A, Dunham WR, Sands RH and Aasa R (1992) Multifrequency EPR investigations into the origin of the S2-state signal at g = 4 of the O2-evolving complex. Biochim Biophys Acta 1099: 25-34

    Article  PubMed  CAS  Google Scholar 

  • Han K-C, Shen J-R, Ikeuchi M and Inoue Y (1994) Chemical cross-linking studies of extrinsic proteins in cyanobacterial Photosystem II. FEBS Lett 355: 121-124

    Article  PubMed  CAS  Google Scholar 

  • Horton RM, Hunt HD, Ho SN, Pullen JK and Pease LR (1989) Engineering hybrid genes without the use of restriction enzymes-gene splicing by overlap extension. Gene 77: 61-68

    Article  PubMed  CAS  Google Scholar 

  • Ioannidis N and Petrouleas V (2000) Electron paramagnetic resonance signals from the S3 state of the oxygen-evolving complex. A broadened radical signal induced by low-temperature near-infrared light illumination. Biochemistry 39: 5246-5254

    Article  PubMed  CAS  Google Scholar 

  • Kirilovsky DL, Boussac AGP, van Mieghem FJE, Ducruet JMRC, Setif PR, Yu JJ, Vermaas WFJ and Rutherford AW (1992) Oxygen evolving Photosystem II preparation from wild-type and Photosystem II mutants of Synechocystis PCC 6803. Biochemistry 31: 2099-2107

    Article  PubMed  CAS  Google Scholar 

  • Kok B, Forbush B and McGloin M (1970) Cooperation of charges in photosynthetic O2 evolution. 1. A linear 4 step mechanism. Photochem Photobiol 11: 457-475

    PubMed  CAS  Google Scholar 

  • Kuhl H, Rogner M, van Breeman JFL and Boekema EJ (1999) Localization of cyanobacterial Photosystem II donor-side subunits by electron microscopy and the supramolecular organization of Photosystem II in the thylakoid membrane. Eur J Biochem 266: 453-459

    Article  PubMed  CAS  Google Scholar 

  • Lakshmi KV, Reifler MJ, Chisholm DA, Wang J, Diner BA and Brudvig GW (2001) Correlation between the g = 4.1 S2 state and the cytochrome c 550 content in cyanobacterial Photosystem II. Biophys J (Annual Meeting Abstracts) 80: 429a-430a

    Google Scholar 

  • Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Meth Enzymol 148: 350-382

    Article  CAS  Google Scholar 

  • Lindberg K and Andréasson L-E (1996) A one-site, two-state model for the binding of anions in Photosystem II. Biochemistry 35: 14259-14267

    Article  PubMed  CAS  Google Scholar 

  • Lorigan GA and Britt RD (2000) Electron spin-lattice relaxation studies of different forms of the S2 state multiline EPR signal of the Photosystem II oxygen-evolving complex. Photosynth Res 66: 189-198

    Article  PubMed  CAS  Google Scholar 

  • MacDonald GM, Boerner RJ, Everly RM, Cramer WA, Debus RJ and Barry BA (1994) Comparison of cytochrome b-559 content in Photosystem II complexes from spinach and Synechocystis Species PCC 6803. Biochemistry 33: 4393-4400

    Article  PubMed  CAS  Google Scholar 

  • Matsukawa T, Mino H, Yoneda D and Kawamori A (1999) Dualmode EPR study of new signals from the S3-state of oxygenevolving complex in Photosystem II. Biochemistry 38: 4072-4077

    Article  PubMed  CAS  Google Scholar 

  • McDermott AE, Yachandra VK, Guiles RD, Cole JL, Dexheimer SL, Britt RD, Sauer K and Klein MP (1988) The state of manganese in the photosynthetic apparatus. 9. Characterization of the manganese O2-evolving complex and the iron quinone acceptor complex in Photosystem II from a thermophilic cyanobacterium by electron paramagnetic resonance and X-ray absorption spectroscopy. Biochemistry 27: 4021-4031

    Article  PubMed  CAS  Google Scholar 

  • Messinger J, Robblee JH, Yu WO, Sauer K, Yachandra VK and Klein MP (1997) The S-0 state of the oxygen-evolving complex in Photosystem II is paramagnetic: detection of EPR multiline signal. J Am Chem Soc 119: 11349-11350

    Article  CAS  Google Scholar 

  • Miller A-F and Brudvig GW (1991) A guide to electron paramagnetic resonance spectroscopy of Photosystem II membranes. Biochim Biophys Acta 1056: 1-18

    PubMed  CAS  Google Scholar 

  • Nishiyama Y, Hayashi H, Watanabe T and Murata N (1994) Photosynthetic oxygen evolution is stabilized by cytochrome c550 against heat inactivation in Synechococcus sp. PCC 7002. Plant Physiol 105: 1313-1319

    Article  PubMed  CAS  Google Scholar 

  • Noren GH, Boerner RJ and Barry BA (1991) EPR characterization of an oxygen-evolving Photosystem II preparation from the transformable cyanobacterium Synechocystis 6803. Biochemistry 30: 3943-3950

    Article  PubMed  CAS  Google Scholar 

  • Ono T, Zimmermann J-L, Inoue Y and Rutherford AW (1986) Electron paramagnetic resonance evidence for a modified S-state transition in chloride-depleted Photosystem II. Biochim. Biophys Acta 851: 193-201

    CAS  Google Scholar 

  • Reifler MJ, Chisholm DA, Wang J, Diner BA and Brudvig GW (1998) Engineering and rapid purification of histidine-tagged Photosystem II from Synechocystis PCC 6803. In: Garab G (ed) Photosynthesis: Mechanisms and Effects, Vol 2, pp 1189-1192. Kluwer Academic Publishers, Dordrecht, The Netherlands

    Google Scholar 

  • Rippka R, Deruelles J, Waterbury JB, Herdman M and Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111: 1-61

    Google Scholar 

  • Rögner M, Nixon PJ and Diner BA (1990) Purification and characterization of Photosystem I and Photosystem II core complexes from wild-type and phycocyanin-deficient strains of the cyanobacterium Synechocystis PCC 6803. J Biol Chem 265: 6189-6196

    PubMed  Google Scholar 

  • Sawaya MR, Krogmann DW, Serag A, Ho KK, Yeates TO and Kerfeld CA (2001) Structures of cytochrome c-549 and cytochrome c6 from the cyanobacterium Arthrospira maxima. Biochemistry 40: 9215-9225

    Article  PubMed  CAS  Google Scholar 

  • Schweitzer RH and Brudvig GW (1997) Fluorescence quenching by chlorophyll cations in Photosystem II. Biochemistry 36: 11351-11359

    Article  PubMed  CAS  Google Scholar 

  • Shen J-R and Inoue Y (1993) Binding and functional properties of two new extrinsic components, cytochrome c-550 and a 12-kDa protein, in cyanobacterial Photosystem II. Biochemistry 32: 1825-1832

    Article  PubMed  CAS  Google Scholar 

  • Shen J-R, Ikeuchi M and Inoue Y (1992) Stoichiometric association of extrinsic cytochrome c550 and 12 kDa protein with a highly purified oxygen-evolving Photosystem II core complex from Synechococcus vulcanus. FEBS Lett 301: 145-149

    Article  PubMed  CAS  Google Scholar 

  • Shen J-R, Burnap RL and Inoue Y (1995a) An independent role of cytochrome c-550 in cyanobacterial photosystem II as revealed by double-deletion mutagenesis of psbO and psbV genes in Synechocystis sp. PCC 6803. Biochemistry 34: 12661-12668

    Article  PubMed  CAS  Google Scholar 

  • Shen J-R, Vermaas W and Inoue Y (1995b) The role of cytochrome c-550 as studied through reverse genetics and mutant characterization in Synechocystis sp. PCC 6803. J Biol Chem 270: 6901-6907

    Article  PubMed  CAS  Google Scholar 

  • Stewart DH, Cua A, Chisholm DA, Diner BA, Bocian DF and Brudvig GW (1998) Identification of histidine 118 in the D1 polypeptide of Photosystem II as the axial ligand to chlorophyll Z. Biochemistry 37: 40040-40046

    Google Scholar 

  • Sugiura M and Inoue Y (1999) Highly purified thermo-stable oxygen-evolving Photosystem II core complex from the thermophilic cyanobacterium Synechococcus elongatus having histagged CP43. Plant Cell Physiol 40: 1219-1231

    PubMed  CAS  Google Scholar 

  • Sugiura M, Inoue Y and Minagawa J (1998) Rapid and discrete isolation of oxygen-evolving His-tagged Photosystem II core complex from Chlamydomonas reinhardtii by Ni2+ affinity column chromatography. FEBS Lett 426: 140-144

    Article  PubMed  CAS  Google Scholar 

  • Tang X-S and Diner BA (1994) Biochemical and spectroscopic characterization of a new oxygen-evolving photosystem core complex from the cyanobacterium Synechocystis PCC 6803. Biochemistry 33: 4594-4603

    Article  PubMed  CAS  Google Scholar 

  • Taylor LA and Rose RE (1988) A correction in the nucleotide sequence of the Tn903 kanamycin resistance determinant in pUC4K. Nucl Acids Res 16: 358

    PubMed  CAS  Google Scholar 

  • Thomas PE, Ryan D and Levin W (1976) An improved staining procedure for the detection of the peroxidase activity of cyto189 chrome P-450 on sodium dodecyl sulfate polyacrylamide gels. Anal Biochem 75: 168-176

    Article  PubMed  CAS  Google Scholar 

  • van Gorkom HJ (1974) Identification of reduced primary electronacceptor of Photosystem II as a bound semiquinone anion. Biochim Biophys Acta 347: 439-442

    Article  PubMed  CAS  Google Scholar 

  • Vrettos JS, Reifler MJ, Kievit O, Lakshmi KV, de Paula JC and Brudvig GW (2001) Factors that determine the unusually low reduction potential of cytochrome c550 in cyanobacterial Photosystem II. J Biol Inorg Chem 6: 708-716

    Article  PubMed  CAS  Google Scholar 

  • Zimmermann J-L and Rutherford AW (1986) Electron paramagnetic resonance properties of the S-2 state of the oxygen-evolving complex of photosystem II. Biochemistry 25: 4609-4615

    Article  CAS  Google Scholar 

  • Zouni A, Witt HT, Kern J, Fromme P, Krauss N, SaengerW and Orth P (2001) Crystal structure of Photosystem II from Synechococcus elongatus at 3.8 Å resolution. Nature 409: 739-743

    Article  PubMed  CAS  Google Scholar 

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Author notes

  1. Michael J. Reifler

    Present address: 454 Corporation, Branford, CT, 06405, USA

Authors and Affiliations

  1. Department of Chemistry, Yale University, New Haven, CT, 06520-8107, USA

    K. V. Lakshmi, Michael J. Reifler & Gary W. Brudvig

  2. Experimental Station, E.I. DuPont de Nemours and Company, Wilmington, DE, 19880-0173, USA

    Dexter A. Chisholm, Jamie Y. Wang & Bruce A. Diner

Authors
  1. K. V. Lakshmi
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  2. Michael J. Reifler
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  3. Dexter A. Chisholm
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  4. Jamie Y. Wang
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  5. Bruce A. Diner
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  6. Gary W. Brudvig
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Correspondence to Gary W. Brudvig.

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Lakshmi, K.V., Reifler, M.J., Chisholm, D.A. et al. Correlation of the cytochrome c 2 content of cyanobacterial Photosystem II with the EPR properties of the oxygen-evolving complex. Photosynthesis Research 72, 175–189 (2002). https://doi.org/10.1023/A:1016140902662

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