Skip to main content
SpringerLink
Log in

The status of cysteine residues in the extrinsic 33 kDa protein of spinach photosystem II complexes

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

Abstract

Two cysteine residues of the extrinsic 33 kDa protein in the oxygen-evolving photosystemII (PS II) complexes were found to exist as cystine residues in situ. The 33 kDa protein, when reduced by 2-mercaptoethanol in either the presence or the absence of 6 M guanidine-HCl (Gdn-HCl), could not rebind with the CaCl2-treated PS II complexes, from which the 33 kDa protein was removed, and evolve any oxygen. Two sulfhydryl (SH) groups of the 33 kDa protein were easily reoxidized to a disulfide (S-S) bond by stirring under aerobic conditions with the concomitant regaining of both the binding ability to the CaCl2-treated PS II complexes and the oxygen-evolving activity.

The molecular conformation of the 33 kDa protein was examined by circular dichroic (CD) spectrometry in the UV regions to reveal that the conformation in the reduced state was completely different from those of the untreated and reoxidized states. The disulfide (S-S) bond of the 33 kDa protein is thus essential to maintain the molecular conformation required to function.

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

CD:

circular dichroism

Chl:

chlorophyll

DMQ:

2,5-dimethyl-p-benzoquinone

DTNB:

5,5′-dithio-bis (2-nitrobenzoic acid)

EDTA:

ethylendiamine-tetraacetic acid

Gdn-HCl:

guanidine-hydrochloric acid

PS II:

photosystem II

SDS:

sodium dodecylsulfate

References

  • AbramowiczDA and DismukesGC (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 

  • AkaboriK, ImaokaA and ToyoshimaY (1984) The role of lipids and 17 kDa protein in enhancing the recovery of O2 evolution in cholate-treated thylakoid membrane. FEBS Lett 173: 36–40

    Google Scholar 

  • ÅkerlundHE and JanssonC (1981) Localization of a 34,000 and a 23,000 Mr polypeptide to the lumenal side of the thylakoid membrane. FEBS Lett 124: 229–232

    Google Scholar 

  • ÅkerlundHE, JanssonC and AnderssonB (1982) Reconstitution of photosynthetic water splitting in inside-out thylakoid vesicles and identification of a participating polypeptide. Biochim Biophys Acta 681: 1–10

    Google Scholar 

  • ArnonDI (1949) Copper enzymes in isolated chloroplasts. Polyphenol oxidase in Beta vulgaris. Plant Physiol 24: 1–15

    Google Scholar 

  • BarrR and CraneFL (1982) Sulfhydryl reagents inhibit electron transport in photosystem II of spinach chloroplasts. Biochim Biophys Acta 681: 139–142

    Google Scholar 

  • BeychekS (1966) Circular dichroism of biological macro-molecules. Science 154: 1288–1299

    Google Scholar 

  • EllmanGL (1958) Tissue sulfhydryl groups. Arch Biochem Biophys 82: 70–77

    Google Scholar 

  • GhanotakisDF, TopperJN, BabcockGT and YocumCF (1984) Water-soluble 17 and 23 kDa polypeptides restore oxygen evolution activity by creating a high affinity binding site for Ca2+ on the oxidizing side of photosystem II. FEBS Lett 170: 169–173

    Google Scholar 

  • InoueY, CroftsAR, Govindjee, MurataN, BengerG and SatohK (eds.) (1983) The oxygen-evolving system of photosynthesis. Academic Press, Tokyo.

    Google Scholar 

  • KessissoglouDP, ButlerWM and PecoraroVL (1987) Characterization of mono- and bi-nuclear manganese (II) shiff-base complexes with metal-bisulfide ligation. Inorg Chem 26: 495–503

    Google Scholar 

  • KuwabaraT and MurataN (1979) Purification and characterization of 33 kDa protein of spinach chloroplasts. Biochim Biophys Acta 581: 228–236

    Google Scholar 

  • KuwabaraT and MurataN (1982a) Inactivation of photosynthetic oxygen evolution and concomitant release of three polypeptides in the photosystem II particles of spinach chloroplasts. Plant Cell Physiol 23: 533–539

    Google Scholar 

  • KuwabaraT and MurataN (1982b) An improved purification method and a further charactization of the 33 kDa protein of spinach chloroplasts. Biochim Biophys Acta 680: 210–215

    Google Scholar 

  • KuwabaraT and MurataN (1983) Quantitative analysis of the inactivation of photosynthetic oxygen evolution and the release of polypeptides and manganese in the photosystem II particles of spinach chloroplasts. Plant Cell Physiol 24: 741–747

    Google Scholar 

  • KuwabaraT and MurataN (1984) Chemical and physicochemical characterization of the proteins involved in the oxygen evolution system. In: SybesmaC (ed.) Advances in Photosynthesis Research Vol. 1, pp 371–374. The Hague: Martinus Nijhoff/Dr W. Junk

    Google Scholar 

  • KuwabaraT, MiyaoM, MurataT and MurataN (1985) The function of 33 kDa protein in photosynthetic oxygen evolution system studied by reconstitution experiments. Biochim Biophys Acta 806: 283–289

    Google Scholar 

  • KuwabaraT, ReddyKJ and ShermanLA (1987) Nucleotide sequence of the gene from the cyanobacterium Anacystis nidulans R22 encoding the Mn-stabilizing protein involved in PS II water oxidation. Proc Natl Acad Sci USA 84: 8230–8234

    Google Scholar 

  • LaemmliUK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • MadsenNB and GurdFRN (1956) The interaction of muscle phosphorylase with pCMB. J. Biol. Chem. 223: 1075–1087

    Google Scholar 

  • MarresCA, VanLoonAPGM, OudshoonR, VanSteegH, GrivellVA and SlaterEC (1985) Nucleotide sequence analysis of the nuclear gene coding for manganese superoxide dismutase of yeast mitochondria, a gene previously assumed to code for the Rieske iron-sulfur protein. Eur J Biochem 147: 153–161

    Google Scholar 

  • MiyaoM and MurataN (1984a) Calcium ions can be substituted for the 24 kDa polypeptide in the photosynthetic oxygen evolution. FEBS Lett 168: 118–120

    Google Scholar 

  • MiyaoM and MurataN (1984b) Role of the 33 kDa polypeptide in preserving Mn in the photosynthetic oxygen-evolution system and its replacement by chloride ions. FEBS Lett 170: 350–354

    Google Scholar 

  • MurataN and MiyaoM (1985) Extrinsic membrane proteins in the photosynthetic oxygenevolving complex. Trends Biochem Sci 10: 122–124

    Google Scholar 

  • Oh-okaH, TanakaS, WadaK, KuwabaraT and MurataN (1986) Complete amino acid sequence of 33 kDa protein isolated from spinach photosystem II particles. FEBS Lett 197: 63–66

    Google Scholar 

  • OnoT and InoueY (1984a) Mn preserving extraction of 33, 24 and 16 kDa proteins from O2-evolving photosystem II particles by divalent salt washing. FEBS Lett 164: 255–260

    Google Scholar 

  • OnoT and InoueY (1984b) Reconstitution of photosynthetic oxygen-evolving activity by rebinding of 33 kDa protein to CaCl2-extracted PS II particles. FEBS Lett 166: 381–384

    Google Scholar 

  • WatanabeA, MinamiE, MuraseM, ShinoharaK and KuwabaraT (1987) Biogenesis of photosystem II complex in spinach chloroplasts. In: BigginsJ (ed.) Progress in Photosynthetic Research Vol. IV pp 629–636. Dordrecht: Martinus Nijhoff/ Dr W. Junk

    Google Scholar 

  • YamamotoT, DoiM, TamuraN and NishimuraM (1981) Release of polypeptides from highly active O2-evolving photosystem 2 preparation by Tris treatment. FEBS Lett 133: 265–268

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, Faculty of Science, Osaka University, Machikaneyama, 560, Toyonaka, Osaka, Japan

    Satoshi Tanaka

  2. Department of Biology, Faculty of Science, Kanazawa University, 920, Marunouchi, Kanazawa, Japan

    Keishiro Wada

Authors
  1. Satoshi Tanaka
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keishiro Wada
    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

Tanaka, S., Wada, K. The status of cysteine residues in the extrinsic 33 kDa protein of spinach photosystem II complexes. Photosynth Res 17, 255–266 (1988). https://doi.org/10.1007/BF00035452

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00035452

Key words

Search

Navigation