Abstract
Inside-out vesicles enriched in photosystem (PS) II activity have been isolated by phase-partitioning in a aqueous two phase system (Åkerlund et al. 1976). We have modified the procedure to obtain a clean separation of the photosystems. Moreover, the PS II vesicles herein obtained were of simple polypeptide composition with retained oxygen evolving activity (Henry, Møller 1981). The necessary modifications entailed:
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(i)
Membrane stacking in the presence of 5–10 mM MgC12 to assure a more efficient stacking compared to that obtained with 150 mM NaCl used in the original procedure. The necessity of stacked membranes for the formation of inside-out vesicles has been demonstrated (Anderson et al. 1978).
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(ii)
Membrane disruption at higher pressures achieved by the use of a French Pressure Cell and repetition of the pressure treatments.
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(iii)
An increase in the number of phase partitioning cycles resulting in a preparation of higher purity.
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(iv)
Optimization of the composition of the phase-partitioning system to produce oxygen evolving PS II vesicles with a minimum number of polypeptides.
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References
Anderson B, Simpson DJ and Hoyer-Hansen G (1978) Carlsberg Res.Commun. 43,77–79.
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© 1984 Springer Science+Business Media Dordrecht
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Henry, L.E.A., Møller, B.L. (1984). Separation of the Photosystems with Retention of their Photochemical Activities. In: Sybesma, C. (eds) Advances in Photosynthesis Research. Advances in Agricultural Biotechnology, vol 3. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-4973-2_45
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DOI: https://doi.org/10.1007/978-94-017-4973-2_45
Publisher Name: Springer, Dordrecht
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