Summary
Cell-free reconstitution systems were used to study in vitro synthesis and membrane integration of the D-2 protein of photosystem II encoded in the chloroplast genome. The D-2 protein was synthesized in vitro from a tobacco psbD gene. A large amount of labelled D-2 was incorporated into broad bean thylakoid membranes that were added to the translation assays either before protein synthesis was started or after translation was finished. Most of the incorporated protein was resistant to a wash with 2 M NaBr or 0.1 M NaOH. The extent of membrane integration of newly formed D-2 was affected neither by using partially extracted thylakoids nor by the addition of a stroma fraction to the expression systems. These observations do not suggest the existence of plastid specific factors involved in the integration of an in vitro synthesized chloroplast encoded thylakoid protein. Possibly, factors in the E. coli and rabbit reticulocyte lysate used for in vitro translation were responsible for preserving an integration-competent conformation of the newly synthesized protein.
Keywords
- Thylakoid Membrane
- Rabbit Reticulocyte Lysate
- Reconstitution Experiment
- Thylakoid Membrane Protein
- Stroma Fraction
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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© 1992 Springer Science+Business Media New York
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Friemann, A., Schwarz, H.J., Hachtel, W. (1992). In Vitro Synthesis and Membrane Integration of the Chloroplast Encoded D-2 Protein of Photosystem II. In: Argyroudi-Akoyunoglou, J.H. (eds) Regulation of Chloroplast Biogenesis. Nato ASI Series, vol 226. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-3366-5_39
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DOI: https://doi.org/10.1007/978-1-4615-3366-5_39
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