Skip to main content
Log in

Effect of trypsin on D1/D2-cytochrom b 559 Photosystem 2 reaction center complex and reaction center from Rhodopseudomonas viridis

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

Abstract

Proteolytic enzyme (trypsin) was used to structurally alter the RCs isolated from plant and bacterium as a way of probing the relation between structure (chromophore-apoprotein interactions) and function (photochemical activity). It was found that neither spectral characteristics (absorption spectrum, the 4th derivative of absorption spectrum) nor photochemical activity (pheophytine photoreduction, P680 photooxidation, etc.) were changed dramatically in D1/D2/cytochrom b 559 PS 2 reaction center complex digested with trypsin. The PS 2 RC treated with trypsin migrates by one green band during electrophoresis with dodecylmaltoside. The peptides with a molecular mass higher than 3–4 kDa were not separated from PS 2 RC. These data indicate that digestion of D1 and D2 proteins does not disturb yet the conformation of peptides or their interactions in so-called ‘core’ of RC and the native state of pigments. In contrast to that, the RC from Rhodopseudomonas viridis treated with enzyme has changed absorption spectrum and lost photochemical activity. The stability of the bacterial RC increased after exchange of LDAO by dodecylmaltoside.

This is a preview of subscription content, log in via an institution to check access.

Access this article

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 a

Cyt:

cytochrome

DPC:

diphenylcarbazide

Dodecylmaltoside:

dodecyl-β-D-maltoside

LDAO:

lauryldimethylamino oxide

Pheo:

pheophytine

PS 2:

Photosystem 2

RC:

reaction center

SiMo:

silicomolybdate

SD:

sodium dodecyl sulfate

References

  • Allakhverdiev SI, Setliková E, Klimov W and Setlik (1987) In photoinhibited Photosystem II particles pheophytin photoreduction remains unimpaired. FEBS Lett 226: 186–190

    Google Scholar 

  • Barber J (1987) Thylakoid membrane organization and function. Biochem Plants 10: 96–100

    Google Scholar 

  • Barber J, Chapman DI and Teffer A (1987) Characterisation of a PS 2 reaction center isolated from the chloroplasts of Pisum sativum. FEBS Lett 220: 67–73

    Google Scholar 

  • Chapman DJ, Gounaris K and Barber J (1988) Electron-transport properties of the isolated D1-D2-cytochrome b-559 Photosystem 2 reaction center. Biochem Biophys Acta 933: 423–431

    Google Scholar 

  • Deisenhofer J and Michel H (1989) The photosynthetic reaction centre from the purple bacterium Rhodopseudomonas viridis. EMBO J 8: 2149–2169

    PubMed  Google Scholar 

  • Drozdova NN, Khotchenkov VP and Krasnovsky AA (1985) Structural changes in the reaction centers of photosynthesizing bacteria Rhodopseudomonas viridis induced by lipase and pronase (in Russian) Biochimii 50: 1213–1219

    Google Scholar 

  • Geiger R, Berzborn RJ, Depka B, Oettmeier W and Trebst A (1987) Site directed antisera to the D2 polypeptide subunit of Photosystem 2. Z Naturforsch 42c: 491–498

    Google Scholar 

  • Ghanotakis DF, dePaule JC, Demetrion DM, Bowlby NR, Petersen J, Babcock DT and Yocum CF (1989) Isolation and characterization of the 47 kDa protein and D1-D2-cytochrome b-559 complex. Biochem Biophys Acta 974: 44–53

    PubMed  Google Scholar 

  • Gounaris K, Chapman DJ and Barber J (1989) Isolation and characterisation of D1/D2-cytochrome b-559 complex from Synechocystis 6803. Biochem Biophys Acta 973: 296–301

    Google Scholar 

  • Irrgang K-D, Renger G and Vater J (1986) Identification of Chl-binding proteins in a PS 2 preparation from spinach. FEBS Lett 204: 67–75

    Article  Google Scholar 

  • Marder JB and Barber J (1989) Probing of apoprotein function in the Photosystem 2 reaction center by proteolytic modification (1989). In: Baltscheffsky M (ed) Current Research in Photosynthesis, Vol I, pp 307–310. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Marder JR, Goloubinoff P and Edelman M (1984) Molecular architecture of the rapidly metabolized 32-kDa protein of Photosystem II. J Biol Chem 259: 3900–3908

    PubMed  Google Scholar 

  • Marder JB, Telfer A and Barber J (1988) The D1 polypeptide subunit of the Photosystem 2 reaction centre has a phosphorylation site at its amino terminus. Biochim Biophys Acta 932: 362–365

    Google Scholar 

  • Mattoo A, Pick U, Hoffman-Falk H and Edelman M (1981) The rapidly metabolized 3200-dalton polypeptide of the chloroplast is the ‘proteinaceous shield’ regulating Photosystem II electron transport and mediating diuron herbicide sensitivity. Proc Natl Acad Sci USA 78: 1572–1576

    PubMed  Google Scholar 

  • Moskalenko AA (1990a) Isolation of stable Photosystem 2 reaction centers (in Russian). Biol membrany 7: 736–741

    Google Scholar 

  • Moskalenko AA (1990b) Isolation of native pigment-protein complexes of plant Photosystem 2 (D1/D2/b-559-complex and D1/D2/b-559/CPa47-complex) by electrophoresis (in Russian). Abstracts of Symposium: ‘Electrophoresis-90’ Riga, USSR

  • Moskalenko AA and Erokhin YuE (1974) Spectral properties and stability of pigment-lipoprotein complex ‘B890’ from Chromatium minutissimum. Studia biophysica 44: 17–32

    Google Scholar 

  • Moskalenko AA and Kuznetsova NYu (1990a) Chlorophyll-protein complexes, polypeptide composition and photochemical activity of the particles enriched in Photosystem 2 during trypsin treatment (in Russian). Biol membrany 7: 487–496

    Google Scholar 

  • Moskalenko AA and Kuznetsova NYu (1990b) Trypsin effect on Photosystem 2 core complex. Photosynthetica 24: 16–21

    Google Scholar 

  • Mullet JE, Arntzen CJ (1981) Identification of a 32–34-kilodalton polypeptides as a herbicide receptor protein in Photosystem 2. Biochim Biophys Acta 635: 236–248

    PubMed  Google Scholar 

  • Nanba O, Saton K (1987) Isolation of a Photosystem 2 reaction center consisting of D1 and D2 polypeptides and cytochrome b-559. Proc Natl Acad Sci USA 84: 109–112

    Google Scholar 

  • Okamura MY, Steiner LA and Feher G (1974) Characterization of reaction centers from photosynthetic bacteria I. Subunit structure of the protein mediating the primary photochemistry in Rhodopseudomonas sphaeroides R-26. Biochemistry 13: 1394–1403

    PubMed  Google Scholar 

  • Peter GF, Machold O and Thornber (1988) Identification and isolation of Photosystem I and Photosystem II pigment-proteins from higher plants. In: Harwood JL and Walton TJ (ed) Plant Membranes-Structure, Assembly and Function, pp 17–31. The Biochemical Society, London

    Google Scholar 

  • Renger G (1976) Studies on the structural and functional organization of system II of photosynthesis. The use of trypsin as a structurally selective inhibitor at the outer surface of the thylakoid membrane. Biochim Biophys Acta 440: 287–300

    PubMed  Google Scholar 

  • Roth M, Lewit-Bentley A, Michel H, Deisenhofer J, Hubert R and Oesterhelt D (1989) Detergent structure in crystals of a bacterial photosynthetic reaction centre. Nature 340: 659–662

    Article  Google Scholar 

  • Rutherford AW (1989) Photosystem II, the water-splitting enzyme. TIBS 14: 227–232

    PubMed  Google Scholar 

  • Sayre RT, Andersson B, Bogorad L (1986) The topology of a membrane protein; The orientation of the 32 kd QB-binding chloroplast thylakoid membrane protein. Cell 47: 601–608

    Article  PubMed  Google Scholar 

  • Schägger H and vonJagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Analit Biochem 166: 368–379

    Google Scholar 

  • Takahashi Y, Satoh K and Itoh S (1989) Silicomolybdate substitutes for the function of a primary electron acceptor and stabilizes charge separation in the Photosystem II reaction center complex. FEBS Lett 255: 133–138

    Article  Google Scholar 

  • Telfer A and Barber J (1989) Evidence for the photoinduced oxidation of the primary electron donor P680 in the isolated Photosystem II reaction centre. FEBS Lett 246: 223–228

    Article  Google Scholar 

  • Tetenkin VL, Gulyaev BA, Seibert M and Rubin AB (1989) Spectral properties of stabilized D1/D2/cytochrome b-559 Photosystem II reaction center complex. Effects of Triton X-100, the redox state of pheophytin and β-carotene. FEBS Lett 250: 459–463

    Article  Google Scholar 

  • Trebst A (1986) The topology of the plastoquinone and herbicide binding peptides of Photosystem II in the thylakoid membrane. Z Naturforsch 41c: 240–245

    Google Scholar 

  • Trebst A and Depka B (1985) The architecture of Photosystem II in Plant Photosynthesis: Which Peptide Subunits carry the Reaction Center of PS II? In: Michel-Beyerle ME (ed) Antennas and Reaction Centers of Photosynthetic Bacteria, pp 216–224. Springer-Verlag, Berlin

    Google Scholar 

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

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moskalenko, A.A., Kuznetsova, N.Y. Effect of trypsin on D1/D2-cytochrom b 559 Photosystem 2 reaction center complex and reaction center from Rhodopseudomonas viridis . Photosynth Res 35, 227–234 (1993). https://doi.org/10.1007/BF00016554

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00016554

Key words

Navigation