Abstract
Superoxide anion radical formation was studied with isolated spinach thylakoid membranes and oxygen evolving Photosystem II sub-thylakoid preparations using the reaction between superoxide and Tiron (1,2-dihydroxybenzene-3,5-disulphonate) which results in the formation of stable, EPR detectable Tiron radicals.
We found that superoxide was produced by illuminated thylakoids but not by Photosystem II preparations. The amount of the radicals was about 70% greater under photoinhibitory conditions than under moderate light intensity. Superoxide production was inhibited by DCMU and enhanced 4–5 times by methyl viologen. These observations suggest that the superoxide in illuminated thylakoids is from the Mehler reaction occurring in Photosystem I, and its formation is not primarily due to electron transport modifications brought about by photoinhibition.
Artificial generation of superoxide from riboflavin accelerated slightly the photoinduced degradation of the Photosystem II reaction centre protein D1 but did not accelerate the loss of oxygen evolution supported by a Photosystem II electron acceptor. However, analysis of the protein breakdown products demonstrated that this added superoxide did not increase the amount of fragments brought about by photoinhibition but introduced an additional pathway of damage.
On the basis of the above observations we propose that superoxide redicals are not the main promoters of acceptor-side-induced photoinhibition of Photosystem II.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- DCBQ-:
-
2,5-dichloro-p-benzoquinone
- DCMU-:
-
3- (3,4-dichlorophenyl)-1,1-dimethylurea
- DMBQ-:
-
2,5-dimethyl-p-benzoquinone
- DMPO-:
-
5,5-dimethyl-pyrrolin N-oxide
- Hepes-:
-
N-(2-hydroxyethyl)-piperazine-N′-(2-ethanesulfonic acid)
- Mes-:
-
2-(N-morpholino)-ethanesulfonic acid
- methyl viologen-:
-
1,1′-dimethyl-4,4′-bipyridinium dichloride
- PS-:
-
Photosystem
- SOD-:
-
Superoxide dismutase (EC 1.15.1.1)
- Tiron-:
-
1,2-dihydroxybenzene-3,5-disulphonate
- Tris-:
-
2-amino-2-hydroxymethylpropane-1,3-diol
References
Ananyev G, Renger G, Wacker U and Klimov V (1994) The photoproduction of superoxide radicals and the superoxide dismutase activity of Photosystem II. The possible involvement of cytochrome b559. Photosynth Res 41: 327–338
Andersson B and Styring S (1991) Photosystem 2-organization, function and acclimation. In: Lee CP (ed) Current Topics in Bioenergetics, Vol 16, pp. 1–81. Academic Press, San Diego
Aro E-M, Virgin I and Andersson B (1993) Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. Biochim Biophys Acta 1143: 113–134
Asada K and Takahashi M (1987) Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CB and Arntzen ChJ (eds) Topics in Photosynthesis, Photoinhibition, Vol 9, pp 227–288. Elsevier, Amsterdam
Barbato R, Friso G, Giardi MT, Rigoni F and Giacometti GM (1991) Breakdown of the Photosystem II reaction centre D1 protein under photoinhibitory conditions: Identification and localization of the C-terminal degradation products. Biochemistry 30: 10220–10226
Barbato R, Friso G, Rigoni F, Frizzo A and Giacometti GM (1992) Characterization of a 41 kDa photoinhibition adduct in isolated Photosystem II reaction centers. FEBS Lett 309: 165–169
Barber J and Andersson B (1992) Too much of a good thing: Light can be bad for photosynthesis. TIBS 17: 61–66
Barényi B and Krause GH (1985) Inhibition of photosynthetic reactions by light. A study with isolated spinach chloroplasts. Planta 163: 218–226
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
Buettner GR and Mason RP (1990) Spin-trapping methods for detecting superoxide and hydroxyl free radicals in vitro and in vivo. In: Packer L and Glazer AN (eds) Meth Enzymol 186, pp 127–133 Academic Press, San Diego
Chen G-X, Kazimir J and Cheniae GM (1992) Photoinhibition of hydroxylamine-extracted Photosystem II membranes: Studies of the mechanism. Biochemistry 31: 11072–11083
Chen G-X, Blubaugh DJ, Homann PH, Golbeck JH and Cheniae GM (1995) Superoxide contributes to the rapid inactivation of specific secondary donors of the Photosystem II reaction center during photodamage of manganese-depleted Photosystem II membranes. Biochemistry 34: 2317–2332
Evans CA (1979) Spin trapping. Aldrichim Acta 12: 23–29
Finkelstein E, Rosen GM and Rauckman EJ (1979) Spin trapping of superoxide. Mol Pharmacol 16: 676–685
Finkelstein E, Rosen GM and Rauckman EJ (1980) Spin trapping. Kinetics of the reaction of superoxide and hydroxylradicals with nitrones. J Am Chem Soc 102: 4994–4999
Ghanotakis DF, Demetriou DM and Yokum CF (1987) Isolation and characterization of an oxygen-evolving Photosystem II reaction center core preparation and a 28 kDa Chl-a binding protein. Biochim Biophys Acta 891: 15–21
Greenstock CL and Miller RW (1975) The oxidation of Tiron by superoxide anion. Kinetics of the reaction in aqueous solutions and chloroplasts. Biochim Biophys Acta 396: 11–16
Grover TA and Piette LH (1981) Influence of flavin addition and removal on the formation of superoxide by NADPH-cytochrome P-450 reductase: A spin-trap study. Arch Biochem Biophys 212: 105–114
Haber F and Weiss JJ (1984) The catalytic decomposition of H2O2 by iron salts. Proc R Soc London (Biol) A147: 332–351
Halliwell B (1978) Superoxide-dependent formation of hydroxyl radicals in the presence of iron chelates. Is it a mechanism for hydroxyl radical production in biochemical systems? FEBS Lett 92: 321–326
Harbour JR and Bolton JR (1975) Superoxide formation in spinach chloroplasts: Electron spin resonance detection by spin trapping. Biochem Biophys Res Commun 64: 803–807.
Herbert SK, Samson G, Fork DC and Laudenbach DE (1992) Characterization of damage to Photosystems I and II in a cyanobacterium lacking detectable iron superoxide dismutase activity. Proc Natl Acad Sci USA 89: 8716–8720
Hideg É and Vass I (1993) The 75°C thermoluminescence band of green tissues: Chemiluminescence from membrane-chlorophyll interaction. Photochem Photobiol 58: 280–283
Hideg É, Spetea C and Vass I (1994a) Singlet oxygen production in thylakoid membranes during photoinhibition as detected by EPR spectroscopy. Photosynth Res 39: 191–199
Hideg É, Spetea C and Vass I (1994b) Singlet oxygen and free radical production during acceptor- and donor-side-induced photoinhibition. Studies with spin trapping EPR spectroscopy. Biochim Biophys Acta 1186: 143–152
Hiramatsu M and Kohno M (1987) Determination of superoxide dismutase activity by electron spin resonance spectroscopy using the spin trap method. JEOL News 23A: 6–9
Janzen EG (1971) Spin trapping. Acc Chem Res 4: 31–39
Kyle DJ (1987) The biochemical basis for photoinhibition of Photosystem II. In: Kyle DJ, Osmond CB and Arntzen ChJ (eds) Topics in Photosynthesis, Photoinhibition, Vol 9, pp 196–225. Elsevier, Amsterdam
Kyle DJ, Ohad P and Arntzen CJ (1984) Membrane protein damage and repair: Selective loss of a quinone-protein function in chloroplast membranes. Proc Natl Acad Sci USA 81: 4070–4074
Macpherson AN, Telfer A, Barber J and Truscott TG (1993) Direct detection of singlet oxygen from isolated photosystem two reaction centers. Biochim Biophys Acta 1143: 301–309
Matheson IBC, Etheridge RD, Kratowich NR and Lee J (1975) The quenching of singlet oxygen by amino acids and proteins. Photochem Photobiol 21: 165–171
McRae DG and Thompson JE (1983) Scenescence-dependent changes in superoxide anion production by illuminated chloroplasts from bean leaves. Planta 158: 185–193
Miller RW and MacDowall FDH (1975) The Tiron free radical as a sensitive indicator of chloroplastic photoautooxidation. Biochim Biophys Acta 387: 176–187
Mishra NP, Mishra RK and Singhal GS (1993) Involvement of active oxygen species in photoinhibition of Photosystem II: Protection of photosynthetic efficiency and inhibition of lipid peroxidation by superoxide dismutase and catalase. J Photochem Photobiol B Biol 19: 19–24
Miyao M (1994) Involvement of active oxygen species in degradation of the D1 protein under strong illumination in isolated subcomplexes of Photosystem II. Biochemistry 33: 9722–9730
Namba O and Satoh K (1987) Isolation of a Photosystem II reaction center consisting of D-1 and D-2 polypeptides and cytochrome b-559. Proc Natl Acad Sci USA 84: 109–112
Powles SB (1984) Photoinhibition of photosynthesis induced by visible light. Annu Rev Plant Physiol 35: 15–44
Prasil O, Adir N and Ohad I (1992) Dynamics of Photosystem II: Mechanism of photoinhibition and recovery processes. In: Barber J (ed) Topics in Photosynthesis, The Photosystems: Structure, Function and Molecular Biology, Vol 11, pp 220–250. Elsevier, Amsterdam
Richter M, Ruhle W and Wild A (1990) Studies on the mechanism of Photosystem II photoinhibition. II. The involvement of toxic oxygen species. Photosynth Res 24: 237–243
Salter AH, Virgin I, Hagman A and Andersson B (1992) On the molecular mechanism of light induced D1 protein degradation in Photosystem II core particles. Biochemistry 31: 3990–3998
Samuni A, Black CDV, Krishna CM, Malech HL, Bernstein EF and Russo A (1988) Hydroxyl radical production by stimulated neutrophils reappraised. J Biol Chem 263: 13797–13801
Sopory SK, Greenberg BM, Mehta RA, Edelman M and Mattoo AK (1990) Free radical scavengers inhibit light-dependent degradation of the 32kDa Photosystem II reaction center protein. Z Naturforsch 45C: 412–417
Styring S, Virgin I, Ehrengberg A and Andersson B (1990) Strong light photoinhibition of electron transport in Photosystem II. Impairment of the function of the first quinone acceptor. Biochim Biophys Acta 1015: 269–278
Takahashi M and Asada K (1982) Dependence of oxygen affinity for Mehler reaction on photochemical activity of chloroplast thylakoids. Plant Cell Physiol 23: 1457–1461
Takahashi Y and Katoh S (1984) Triplet states in Photosystem I reaction center complex. Inhibition of radical pair recombination by bipyridinium dyes and naphtoquinones. Plant Cell Physiol 25: 785–794
Telfer A, Drami S, Bishop S M, Philips D and Barber J (1994) β-carotene quenches singlet oxygen formed by isolated Photosystem II reaction centers. Biochemistry 33: 14469–14474
Tschiersch H and Ohmann E (1993) Photoinhibition in Euglena gracilis: Involvement of active oxygen species. Planta 191: 316–332
Van Ginkel G and Raison JK (1980) Light-induced formation of O2 − oxygen radicals in systems containing chlorophyll. Photochem Photobiol 32: 793–798
Van Mieghem FJE, Nitschke W, Mathis P and Rutherford AW (1989) The influence of the quinone-iron electron acceptor complex on the reaction centre photochemistry of Photosystem II. Biochim Biophys Acta 977: 207–214
Vass I and Styring S (1992) Spectroscopic characterization of triplet forming states in Photosystem II. Bichemistry 31: 5957–5693
Vass I and Styring S (1993) Characterization of chlorophyll triplet promoting states in Photosystem II sequentially induced during photoinhibition. Biochemistry 32: 3334–3341
Vass I, Styring S, Hundall T, Koivuniemi A, Aro E-M and Andersson B (1992) Reversible and irreversible intermediates during photoinhibition of Photosystem 2. Stable reduced QA species promote chlorophyll triplet formation. Proc Natl Acad Sci USA 89: 1408–1412
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hideg, É., Spetea, C. & Vass, I. Superoxide radicals are not the main promoters of acceptor-side-induced photoinhibitory damage in spinach thylakoids. Photosynth Res 46, 399–407 (1995). https://doi.org/10.1007/BF00032294
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00032294