Photosynthetica 2002, 40(3):355-361 | DOI: 10.1023/A:1022666706700
Isolation and Characterization of an Oxygen Evolving Photosystem 2 Core Complex from the Thermophilic Cyanobacterium Mastigocladus laminosus
- 1 Department of Botany, The Hebrew University of Jerusalem, Givat Ram, Jerusalem, Israel
A novel purification procedure was developed for the isolation of oxygen evolving photosystem 2 (PS2) from Mastigocladus laminosus. The isolation procedure involves dodecyl maltoside extraction followed by column chromatography using anion exchange resins. The isolated PS2 reaction center (RC) was analyzed for its biochemical and biophysical characteristics. Analysis by SDS polyacrylamide gel electrophoresis revealed that the complex contained five intrinsic membrane proteins (CP 47, CP 43, D1, D2, and cyt b559) and at least three low molecular mass proteins. The complex exhibited high rates of oxygen evolution [333 mmol(O2) kg-1(Chl) s-1] in the presence of 2.5 mM 2,6-dimethylbenzoquinone (DMBQ) as an artificial electron acceptor. The red chlorophyll a absorption peak of this complex was observed at 673.5±0.2 nm. The isolated PS2 core complex was free of photosystem 1 as inferred from its SDS-PAGE and fluorescence spectrum. The electron transfer properties of the Mastigocladus cells and the purified PS2 core complex were further probed by measuring thermoluminescence signals, which indicated the presence of a primary quinone electron acceptor (QA) in the purified PS2 core complex.
Additional key words: 2,6-dimethylbenzoquinone; fluorescence; membrane proteins; oxygen evolution rate; reaction center; thermoluminescence
Published: September 1, 2002 Show citation
| ACS | AIP | APA | ASA | Harvard | Chicago | Chicago Notes | IEEE | ISO690 | MLA | NLM | Turabian | Vancouver |
References
- Arnon, D.I., McSwain, B.D., Tsujimoto, H.Y., Wada, K.: Photochemical activity and components of membrane preparations from blue-green algae. I. Coexistence of two photosystems in relation to chlorophyll a and removal of phycocyanin.-Biochim. biophys. Acta 357: 231-245, 1974.
Go to original source... - Bianchetti, M., Zheleva, D., Deak, Z., Zharmuhamedov, S., Klimov, V., Nugent, J., Vass, I., Barber, J.: Comparison of the functional properties of the monomeric and dimeric forms of the isolated CP47-reaction center complex.-J. biol. Chem. 273: 16128-16133, 1998.
Go to original source... - Binder, A., Bachofen, R.: Isolation and characterization of a coupling factor I ATPase of the thermophilic blue-green alga (cyanobacterium) Mastigocladus laminosus.-FEBS Lett. 104: 66-70, 1979.
Go to original source... - Bricker, T.M., Ghanotakis, D.F.: Introduction to oxygen evolution and the oxygen-evolving complex.-In: Ort, D.R., Yocum, C.F. (ed.): Oxygenic Photosynthesis: The Light Reactions. Pp. 113-136. Kluwer Academic Publishers, Dordrecht-Boston-London 1996.
Go to original source... - Bumann, D., Oesterhelt, D.: Purification and characterization of oxygen-evolving photosystem II core complexes from the green alga Chlamydomonas reinhardtii.-Biochemistry 33: 10906-10910, 1994.
Go to original source... - Castenholz, R.W.: Thermophilic blue-green algae and the thermal environment.-Bacteriol. Rev. 33: 476-504, 1969.
Go to original source... - Debus, R.J.: The manganese and calcium ions of photosynthetic oxygen evolution.-Biochim. biophys. Acta 1102: 269-352, 1992.
Go to original source... - Gantt, E.: Pigment protein complexes and the concept of the photosynthetic unit: Chlorophyll complexes and phycobilisomes.-Photosynth. Res. 48: 47-53, 1996.
Go to original source... - Gounaris, K., Chapman, D.J., Booth, P., Crystall, B., Giorgi, L.B., Klug, D.R., Porter, G., Barber, J.: Comparison of the D1/D2 cytochrome b-559 reaction centre complex of photosystem two isolated by two different methods.-FEBS Lett. 265: 88-92, 1990.
Go to original source... - Hankamer, B., Nield, J., Zheleva, D., Boekema, E., Jansson, S., Barber, J.: Isolation and biochemical characterisation of monomeric and dimeric photosystem II complexes from spinach and their relevance to the organization of photosystem II in vivo.-Eur. J. Biochem. 243: 422-429, 1997.
Go to original source... - Ikeuchi, M., Inoue, Y.: Characterization of O2 evolution by a wheat photosystem II reaction center complex isolated by a simplified method: disjunction of secondary acceptor quinone and enhanced Ca2+ demand.-Arch. Biochem. Biophys. 247: 97-107, 1986.
Go to original source... - Inoue, Y.: Photosynthetic thermoluminescence as a simple probe of photosystem II electron transport.-In: Amesz, J., Hoff, A.J. (ed.): Biophysical Techniques in Photosynthesis. Pp. 93-107. Kluwer Academic Publishers, Dordrecht-Boston-London 1996.
Go to original source... - Keren, N., Berg, A., van Kan, P.J., Levanon, H., Ohad, I.: Mechanism of photosystem II photoinactivation and D1 protein degradation at low light: The role of back electron flow.-Proc. nat. Acad. Sci. USA 94: 1579-1584, 1997.
Go to original source... - Kirilovsky, D.L., Boussac, A.G.P., Van Mieghem, F.J.E., Ducruet, J.-M.R.C., Sétif, P.R., Yu, J., Vermaas, W.F.J., Rutherfold, A.W.: Oxygen-evolving photosystem II preparation from wild type and photosystem II mutants of Synechocystis sp. PCC 6803.-Biochemistry 31: 2099-2107, 1992.
Go to original source... - Koike, H., Mamada, K., Ikeuchi, M., Inoue, Y.: Low-molecular-mass proteins in cyanobacterial photosystem II: identification of psbH and psbK gene products by N-terminal sequencing.-FEBS Lett. 244: 391-396, 1989.
Go to original source... - MacDonald, G.M., Barry, B.A.: Difference FT-IR study of a novel biochemical preparation of photosystem II.-Biochemistry 31: 9848-9856, 1992.
Go to original source... - Nechushtai, R., Muster, P., Binder, A., Liveanu, V., Nelson, N.: Photosystem I reaction center from the thermophilic cyanobacterium Mastigocladus laminosus.-Proc. nat. Acad. Sci. USA 80: 1179-1183, 1983.
Go to original source... - Noren, G.H., Boerner, R.J., Barry, B.A.: EPR characterization of an oxygen-evolving photosystem-II preparation from the transformable cyanobacterium Synechocystis 6803.-Biochemistry 30: 3943-3950, 1991.
Go to original source... - Oren-Shamir, M., Sai, P.S.M., Edelman, M., Scherz, A.: Isolation and spectroscopic characterization of a plantlike photosystem II reaction center from the cyanobacterium Synechocystis sp. 6803.-Biochemistry 34: 5523-5526, 1995.
Go to original source... - Rögner, M., Nixon, P.J., Diner, B.A.: 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, 1990.
Go to original source... - Schagger, H., Aquila, H., Jagow, G. von: Coomassie blue-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for direct visualization of polypeptides during electrophoresis.-Anal. Biochem. 173: 201-205, 1988.
Go to original source... - Stewart, A.C., Bendall, D.S.: Photosynthetic electron transport in a cell-free preparation from the thermophilic blue-green alga Phormidium laminosum.-Biochem. J. 188: 351-361, 1980.
Go to original source... - Tang, X.-S., Diner, B.A.: Biochemical and spectroscopic characterization of a new oxygen-evolving photosystem II core complex from the cyanobacterium Synechocystis PCC 6803.-Biochemistry 33: 4594-4603, 1994.
Go to original source... - Tang, X.-S., Fushimi, K., Satoh, K.: D1-D2 complex of the photosystem II reaction center from spinach. Isolation and partial characterization.-FEBS Lett. 273: 257-260, 1990.
Go to original source... - Zer, H., Prasil, O., Ohad, I.: Role of plastoquinol oxidoreduction in regulation of photochemical reaction center II D1 protein turnover in vivo.-J. biol. Chem. 269: 17670-17676, 1994.
Go to original source...




