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Investigation of the spatial relationships between photosystem 2 polypeptides by reversible crosslinking and diagonal electrophoresis

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Abstract

Nearest neighbour relationships within the LHC2-PS2 complex were investigated by using the reversible crosslinking agent dithiobis(succinimidyl propionate) (DSP). This was accomplished by treating PS2-enriched membranes, prepared from chloroplasts of Pisum sativum, with the crosslinker followed by diagonal electrophoresis of the solubilised polypeptides.

Analysis of the off-diagonal spot patterns produced by crosslinker cleavage and second dimension electrophoresis was made on the basis of: staining with Coomassie blue or silver, labelling with [35S]-methionine, and sensitivity to 1 M NaCl washing. It was concluded that LHC2 polypeptides crosslinked with several components of the PS2 complex and that the extrinsic polypeptides associated with water oxidation, having approximate molecular weights of 16 and 23 kDa, crosslink to form homodimers. The latter finding suggests that there may be more than one copy of each of these polypeptides per PS2 complex.

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Abbreviations

DMSO:

dimethylsulphoxide

DSP:

dithiobis(succinimidylpropionate)

DCPIP:

dichlorophenolindophenol

SDS:

sodium dodecylsulphate

PS2:

photosystem 2

LHC2:

light harvesting chlorophyll a/b complex associated with photosystem 2

MES:

2[N-morpholino] ethanesulphonic acid

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Author notes

  1. P. A. Millner

    Present address: Department of Biochemistry, Leeds University, Leeds, UK

  2. G. Gogel

    Present address: Department of Chemistry, Colgate University, 13346, Hamilton, N.Y., USA

Authors and Affiliations

  1. AFRC Photosynthesis Research Group, Department of Pure and Applied Biology, Imperial College of Science and Technology, SW7 2BB, London, UK

    P. A. Millner, G. Gogel & J. Barber

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  1. P. A. Millner
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  2. G. Gogel
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  3. J. Barber
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Millner, P.A., Gogel, G. & Barber, J. Investigation of the spatial relationships between photosystem 2 polypeptides by reversible crosslinking and diagonal electrophoresis. Photosynth Res 13, 185–198 (1987). https://doi.org/10.1007/BF00035240

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  • DOI: https://doi.org/10.1007/BF00035240

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