In Vitro reconstitution of synthesis, uptake, and assembly of cytoplasmically synthesized chloroplast proteins
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The process by which nuclear coded polypeptides are assembled into the chloroplast has been reconstituted in vitro and involves three sequential events: (1) translation of cytoplasmic RNA; (2) uptake of translation products into the chloroplasts; and (3) sorting and assembly of polypeptides to sites in the envelope, stroma, and thylakoid membrane. Translation products imported by chloroplasts are sorted to the envelope membrane, stroma, and the thylakoid membrane. A technique to isolate chloroplast envelope from intact chloroplasts is described in this chapter. The chloroplast thylakoid membrane contains four major protein complexes: adenosine triphosphatase (ATPase), Cyt b6/f complex, photosystem I, and photosystem II. Each of these complexes consists of polypeptides synthesized within the chloroplast and polypeptides synthesized on cytoplasmic ribosomes. The isolation of a thylakoid membrane complex following the uptake of 35S-labeled translation products provides a method to test for assembly of the newly imported polypeptides and to determine the site of synthesis of polypeptides in a given protein complex. © 1983, Elsevier Inc. All rights reserved.
author list (cited authors)
Mullet, J. E., & Chua, N.