Photosynthetica 2005, 43(3):421-424 | DOI: 10.1007/s11099-005-0067-1

Secondary structure estimation of recombinant psbH, encoding a photosynthetic membrane protein of cyanobacterium Synechocystis sp. PCC 6803

D. Stys1,2, W. Schoefberger1,2, Z. Halbhuber1,2, J. Ristvejova1,2, N. Muller3, R. Ettrich1,2,*
1 Laboratories of High Performance Computing and Biomembranes, Institute of Physical Biology, University of South Bohemia, Nove Hrady, Czech Republic
2 Institute of Systems Biology and Ecology, Academy of Sciences of the Czech Republic, Nove Hrady, Czech Republic
3 Johannes Kepler Universitat, Linz, Austria

The PsbH protein of cyanobacterium Synechocystis sp. PCC 6803 was expressed as a fusion protein with glutathione-S transferase (GST) in E. coli grown on a mineral medium enriched in 15N isotope. After enzymatic cleavage of the fusion protein, the 1H-15N-HSQC spectrum of PsbH protein in presence of the detergent β-D-octyl-glucopyranoside (OG) was recorded on a Bruker DRX 500 MHz NMR spectrometer equipped with a 5 mm TXI cryoprobe to enhance the sensitivity and resolution. Non-labelled protein was used for secondary structure estimation by deconvolution from circular dichroism (CD) spectra. Experimental results were compared with our results from a structural model of PsbH using a restraint-based comparative modelling approach combined with molecular dynamics and energetic modelling. We found that PsbH shows 34-38% α-helical structure (Thr36-Ser60), a maximum of around 15% of β-sheet, and 12-19% of β-turn.

Additional key words: CD spectroscopy; molecular dynamics calculations; NMR spectroscopy; photosystem 2; protein folding

Received: February 22, 2005; Accepted: May 23, 2005; Published: September 1, 2005  Show citation

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Stys, D., Schoefberger, W., Halbhuber, Z., Ristvejova, J., Muller, N., & Ettrich, R. (2005). Secondary structure estimation of recombinant psbH, encoding a photosynthetic membrane protein of cyanobacterium Synechocystis sp. PCC 6803. Photosynthetica, 43(3), 421-424. doi: 10.1007/s11099-005-0067-1
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