Skip to main content
SpringerLink
Log in

Formation of the S2 state and structure of the Mn complex in photosystem II lacking the extrinsic 33 kilodalton polypeptide

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

Abstract

Electron paramagnetic resonance (EPR) spectroscopy and O2 evolution assays were performed on photosystem II (PSII) membranes which had been treated with 1 M CaCl2 to release the 17, 23 and 33 kilodalton (kDa) extrinsic polypeptides. Manganese was not released from PSII membranes by this treatment as long as a high concentration of chloride was maintained. We have quantitated the EPR signals of the several electron donors and acceptors of PSII that are photooxidized or reduced in a single stable charge separation over the temperature range of 77 to 240 K. The behavior of the samples was qualitatively similar to that observed in samples depleted of only the 17 and 23 kDa polypeptides (de Paula et al. (1986) Biochemistry25, 6487–6494). In both cases, the S2 state multiline EPR signal was observed in high yield and its formation required bound Ca2+. The lineshape of the S2 state multiline EPR signal and the magnetic properties of the manganese site were virtually identical to those of untreated PSII membranes. These results suggest that the structure of the manganese site is unaffected by removal of the 33 kDa polypeptide. Nevertheless, in samples lacking the 33 kDa polypeptide a stable charge separation could only be produced in about one half of the reaction centers below 160 K, in contrast to the result obtained in untreated or 17 and 23 kDa polypeptide-depleted PSII membranes. This suggests that one function of the 33 kDa polypeptide is to stabilize conformations of PSII that are active in secondary electron transfer events.

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

Abbreviations

Chl-:

chlorophyll

DCBQ-:

2,5-dichloro-p-benzoquinone

DCMU-:

(diuron) 3-(3,4-dichlorophenyl)-1,1-dimethylurea

EGTA-:

ethylene glycol bis-(β-aminoethyl ether) N,N,N′,N′-tetraacetic acid

EPR-:

electron paramagnetic resonance

HSB-:

high salt buffer

HSCaB-:

high salt Ca2+ buffer

kDa-:

kilodalton

MES-:

2-(N-morpholino)ethanesulfonic acid

P680-:

primary electron donor in PSII

PaGE-:

polyacrylamide gel electrophoresis

PSII-:

Photosystem II

QA-:

primary quinone electron acceptor in PSII

RB-:

resuspension buffer

TMPD-:

N,N,N′,N′-tetramethyl-p- phenylenediamine

Tris-:

tris(hydroxymethyl)aminomethane

TX100-:

Triton X-100

Z-:

endogenous electron donor to P680+

References

  1. Abramowicz DA and Dismukes GC (1984) Manganese proteins isolated from spinach thylakoid membranes and their role in O2 evolution. II. A binuclear manganese-containing 34 kilodalton protein, a probable component of the water dehydrogenase enzyme. Biochim Biophys Acta 765:318–328

    Google Scholar 

  2. Andersson B, Ljungberg U, Åkerlund H-E and Bishop N (1985) Discrimination by immunological analysis between two 33–34 kDa polypeptides involved in photosynthetic oxygen evolution. Biochim Biophys Acta 809:288–290

    Google Scholar 

  3. Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenol oxidase inBeta vulgaris. Plant Physiol 24:1–15

    Google Scholar 

  4. Babcock GT, Ghanotakis DF, Ke B and Diner BA (1983) Electron donation to photosystem II in reaction center preparations. Biochim Biophys Acta 723:276–286

    Google Scholar 

  5. Beck WF, dePaula JC and Brudvig GW (1985) Active and resting states of the O2-evolving complex of photosystem II. Biochemistry 24:3035–3043

    Google Scholar 

  6. Berthold DA, Babcock GT and Yocum CF (1981) A highly resolved, oxygen-evolving photosystem II preparation from spinach thylakoid membranes. FEBS Lett 134:231–234

    Google Scholar 

  7. Bishop NI (1983) Identification of photosystem II-thylakoid proteins through mutational analysis withChlamydomonas andScenedesmus phenotypes. In Inoue Y, Crofts AR, Murata N, Renger G and Satoh K (eds) The oxygen evolving system of photosynthesis, pp. 177–187. Tokyo: Academic Press

    Google Scholar 

  8. Blough NV and Sauer K (1984) The effects of mono- and divalent salts on the O2-evolution activity and low temperature multiline EPR spectrum of photosystem II preparations from spinach. Biochim Biophys Acta 767:377–381

    Google Scholar 

  9. Chua N-H (1980) Electrophoretic analysis of chloroplast proteins. Methods in Enzymology 69:434–446

    Google Scholar 

  10. Dekker JP, Ghanotakis DF, Plijter JJ, vanGorkom HJ and Babcock GT (1984) Kinetics of the oxygen-evolving complex in salt-washed photosystem II preparations. Biochim Biophys Acta 767:515–523

    Google Scholar 

  11. dePaula JC, Innes JB and Brudvig GW (1985) Electron transfer in photosystem II at cryogenic temperatures. Biochemistry 24:8114–8120

    Google Scholar 

  12. dePaula JC, Beck WF and Brudvig GW (1986) Magnetic properties of manganese in the photosynthetic O2-evolving complex. 2. Evidence for a manganese tetramer. J Am Chem Soc 108:4002–4009

    Google Scholar 

  13. dePaula JC, Li PM, Miller A-F, Wu BW and Brudvig GW (1986) The effect of the 17 and 23 kDa polypeptides, calcium, and chloride on electron transfer in photosystem II. Biochemistry 25:6487–6494

    Google Scholar 

  14. Dismukes GC (1986) The metal centers of the photosynthetic oxygen-evolving complex. Photochem Photobiol 43:99–115

    Google Scholar 

  15. Franzén L-G, Hansson Ö and Andréasson L-E (1985) The roles of the extrinsic subunits in photosystem II as revealed by EPR. Biochim Biophys Acta 808:171–179

    Google Scholar 

  16. Ghanotakis DF, Babcock GT and Yocum CF (1985) Structure of the oxygen-evolving complex of photosystem II: calcium and lanthanum compete for sites on the oxidizing side of photosystem II which control the binding of water-soluble polypeptides and regulate the activity of the manganese complex. Biochim Biophys Acta 809:173–180

    Google Scholar 

  17. Ghanotakis DF and Yocum CF (1985) Polypeptides of photosystem II and their role in oxygen evolution. A minireview. Photosyn Res 7:97–114

    Google Scholar 

  18. Ikeuchi M, Yuasa M and Inoue Y (1985) Simple and discrete isolation of an O2-evolving PSII reaction center complex retaining Mn and the extrinsic 33 kDa protein. FEBS Lett 185:316–322

    Google Scholar 

  19. Imaoka A, Akabori K, Yanagi M, Izumi K, Toyoshima Y, Kawamori A, Nakayama H and Sato J (1986) Roles of the three lumen-surface proteins in the formation of S2 state and O2 evolution in photosystem II particles from spinach thylakoid membranes. Biochim Biophys Acta 848:201–211

    Google Scholar 

  20. Kleinfeld D, Okamura MY and 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

    Google Scholar 

  21. Kuwabara T, Miyao M, Murata T and Murata N (1985) The function of 33-kDa protein in the photosynthetic oxygen-evolution system studied by reconstitution experiments. Biochim Biophys Acta 806:283–289

    Google Scholar 

  22. Matsuura K and Itoh S (1985) Effects of CaCl2-washing on EPR signals of cytochrome b559 in photosystem II preparation from spinach chloroplasts. Plant Cell Physiol 26: 537–542

    Google Scholar 

  23. Metz J, Bricker T and Seibert M (1985) The azido[14C]atrazine photoaffinity technique labels a 34-kDa protein inScenedesmus which functions on the oxidizing side of photosystem II. FEBS Lett 185:191–196

    Google Scholar 

  24. Miyao M and Murata N (1984) Role of the 33-kDa polypeptide in preserving Mn in the photosynthetic oxygen-evolving system and its replacement by chloride ions. FEBS Lett 170:350–354

    Google Scholar 

  25. Miyao M and Murata N (1985) The chloride effect on photosynthetic oxygen evolution: interaction of Cl− with 18 kDa, 24 kDa and 33 kDa proteins. FEBS Lett 180:303–308

    Google Scholar 

  26. Miyao M, Murata N, Maison-Peteri B, Boussac A, Etienne A-L and Lavorel J (1987) Effect of the 33-kDa protein on the S-state transition in the oxygen-evolving complex. In: Biggins J (ed.) Progress in Photosynthesis Research 1:613–616, Dordrecht: Martinus Nijhoff Publishers

    Google Scholar 

  27. Ono T and Inoue Y (1983) Mn-preserving extraction of 33-, 24- and 16-kDa proteins from O2-evolving PS II particles by divalent salt-washing. FEBS Lett 164:255–260

    Google Scholar 

  28. Ono T and Inoue Y (1984) Ca2+-dependent restoration of O2-evolving activity in CaCl2-washed PS II particles depleted of 33, 24 and 16 kDa proteins. FEBS Lett 168:281–286

    Google Scholar 

  29. Ono T and Inoue Y (1985) S-state turnover in the O2-evolving system of CaCl2-washed photosystem II particles depleted of the three peripheral proteins as measured by thermoluminescence. Removal of the 33 kDa polypeptide inhibits S3 to S4 transition. Biochim Biophys Acta 806:331–340

    Google Scholar 

  30. Satoh K and Katoh S (1985) A functional site of Ca2+ in the oxygen-evolving photosystem II preparation fromSynechococcus sp. FEBS Lett 190:199–203

    Google Scholar 

  31. Sétif P, Mathis P and Vänngård T (1984) Photosystem I Photochemistry at low temperature. Heterogeneity in pathways for electron transfer to the secondary acceptors and for recombination processes. Biochim Biophys Acta 767:404–414

    Google Scholar 

  32. Styring S, Miyao M and Rutherford AW (1987) Formation and flash-dependent oscillation of the S2-state multiline EPR-signal in an oxygen evolving PSII-preparation lacking the three extrinsic proteins in the oxygen evolving system. Biochim Biophys Acta 890:32–38

    Google Scholar 

  33. Tamura N, Radmer R, Lantz S, Cammarata K and Cheniae G (1986) Depletion of photosystem II-extrinsic proteins. II. Analysis of the PSII/water-oxidizing complex by measurements of N,N,N′,N′-tetramethyl-p-phenylenediamine oxidation following an actinic flash. Biochim Biophys Acta 850:369–379

    Google Scholar 

  34. Völker M, Ono T, Inoue Y and Renger G (1985) Effect of trypsin on PS-II particles. Correlation between Hill-activity, Mn-abundance and peptide pattern. Biochim Biophys Acta 806:25–34

    Google Scholar 

  35. Yamamoto Y, Shinkai H, Isogai Y, Matsuura K and Nishimura M (1984) Isolation of an Mn-carrying 33-kDa protein from an oxygen-evolving photosystem-II preparation by phase partitioning with butanol. FEBS Lett 175:429–432

    Google Scholar 

  36. Yocum CF, Yerkes CT, Blankenship RE, Sharp RR and Babcock GT (1981) Stoichiometry, inhibitor sensitivity, and organization of manganese associated with photosynthetic oxygen evolution. Proc Natl Acad Sci USA 78:7507–7511

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry, Yale University, 06511, New Haven, CT, USA

    Anne-Frances Miller, Julio C. de Paula & Gary W. Brudvig

Authors
  1. Anne-Frances Miller
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Julio C. de Paula
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Gary W. Brudvig
    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

Miller, AF., de Paula, J.C. & Brudvig, G.W. Formation of the S2 state and structure of the Mn complex in photosystem II lacking the extrinsic 33 kilodalton polypeptide. Photosynth Res 12, 205–218 (1987). https://doi.org/10.1007/BF00055121

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00055121

Key words

Search

Navigation