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J. Biol. Chem., Vol. 281, Issue 43, 32705-32713, October 27, 2006

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Cyanobacterial Small Chlorophyll-binding Protein ScpD (HliB) Is Located on the Periphery of Photosystem II in the Vicinity of PsbH and CP47 Subunits^*

Kamoltip Promnares^¶, Josef Komenda^¶, Ladislav Bumba^||**, Jana Nebesarova^||, Frantisek Vacha^||, and Martin Tichy^¶1

From the Faculty of Biological Sciences, University of South Bohemia, 370 05 Ceske Budejovice, Czech Republic, the Institute of Physical Biology, University of South Bohemia, 373 33 Nové Hrady, Czech Republic, the ^¶Laboratory of Photosynthesis, Institute of Microbiology, Czech Academy of Sciences, 379 81 Trebon, Czech Republic, the ^||Biological Center, Czech Academy of Sciences, 370 05 Ceske Budejovice, Czech Republic, and the ^**Institute of Microbiology, Czech Academy of Sciences, 142 20 Prague, Czech Republic

Cyanobacteria contain several genes coding for small one-helix proteins called SCPs or HLIPs with significant sequence similarity to chlorophyll a/b-binding proteins. To localize one of these proteins, ScpD, in the cells of the cyanobacterium Synechocystis sp. PCC 6803, we constructed several mutants in which ScpD was expressed as a His-tagged protein (ScpDHis). Using two-dimensional native-SDS electrophoresis of thylakoid membranes or isolated Photosystem II (PSII), we determined that after high-light treatment most of the ScpDHis protein in a cell is associated with PSII. The ScpDHis protein was present in both monomeric and dimeric PSII core complexes and also in the core subcomplex lacking CP43. However, the association with PSII was abolished in the mutant lacking the PSII subunit PsbH. In a PSII mutant lacking cytochrome b₅₅₉, which does not accumulate PSII, ScpDHis is associated with CP47. The interaction of ScpDHis with PsbH and CP47 was further confirmed by electron microscopy of PSII labeled with Ni-NTA Nanogold. Single particle image analysis identified the location of the labeled ScpDHis at the periphery of the PSII core complex in the vicinity of the PsbH and CP47. Because of the fact that ScpDHis did not form any large structures bound to PSII and because of its accumulation in PSII subcomplexes containing CP47 and PsbH we suggest that ScpD is involved in a process of PSII assembly/repair during the turnover of pigment-binding proteins, particularly CP47.

Received for publication, July 5, 2006 , and in revised form, August 18, 2006.

^* The work was supported in part by the project MSM6007665808 of the Ministry of Education, Youth and Sports of the Czech Republic, and by the Czech Academy of Sciences Institutional Research Concepts AV0Z50200510 and AV0Z50510513. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: Inst. of Microbiology, Laboratory of Photosynthesis, Opatovicky mlyn, 379 81 Trebon, Czech Republic. Tel.: 420-384-722-268; Fax: 420-384-721-246; E-mail: tichym{at}alga.cz.

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