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Precursors of one integral and five lumenal thylakoid proteins are imported by isolated pea and barley thylakoids: optimisation of in vitro assays

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Abstract

In vitro assays for the import of proteins by isolated pea thylakoids have been refined and optimised with respect to (a) the method of thylakoid preparation, (b) the concentration of thylakoids in the import assay, and (c) the pH and temperature of the import assay. As a result, the 23 kDa and 16 kDa proteins of the photosynthetic oxygen-evolving complex are imported with efficiencies approaching 100%; import of the third oxygen-evolving complex protein is also observed, albeit with lower efficiencies. We have also demonstrated import of three further thylakoid proteins: plastocyanin, the CFoII subunit of the ATP synthase, and the photosystem I subunit, PSI-N, using this import assay. Import of plastocyanin, PSI-N and the 33 kDa oxygen-evolving complex protein subunit requires the presence of stromal extract whereas the other three proteins are efficiently imported in the absence of added soluble proteins. Import into isolated barley thylakoids was achieved under identical assay conditions, although with somewhat lower efficiency than into pea thylakoids.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Warwick, CV4 7AL, Coventry, UK

    Ian Wallace Brock, Laurence Hazell & Colin Robinson

  2. Botanisches Institut der Ludwig-Maximilians-Universität, Menzinger Strasse 67, 80638, München, Germany

    Doris Michl, Reinhold G. Herrmann & Ralf Bernd Klösgen

  3. Plant Biochemistry Laboratory, Department of Plant Biology, Royal Veterinary and Agricultural University, Thorvaldsenvej 40, DK-1871, Frederiksberg C, Denmark

    Vibeke Skovgaard Nielsen, Birger Lindberg Møller & Colin Robinson

Authors
  1. Ian Wallace Brock
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  2. Laurence Hazell
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  3. Doris Michl
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  4. Vibeke Skovgaard Nielsen
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  5. Birger Lindberg Møller
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  6. Reinhold G. Herrmann
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  7. Ralf Bernd Klösgen
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  8. Colin Robinson
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Brock, I.W., Hazell, L., Michl, D. et al. Precursors of one integral and five lumenal thylakoid proteins are imported by isolated pea and barley thylakoids: optimisation of in vitro assays. Plant Mol Biol 23, 717–725 (1993). https://doi.org/10.1007/BF00021527

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  • DOI: https://doi.org/10.1007/BF00021527

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