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Updated: Apr 22, 2026

Studying Protein Import into Chloroplasts Using Protoplasts
Published on: December 10, 2018
Assembly of two functionally-distinct protein import complexes in the outer membrane of plant chloroplasts
Sreedhar Nellaepalli1, Domagoj Baretić2, Astrid F Brandner3
1Section of Molecular Plant Biology, Department of Biology, University of Oxford, Oxford, UK. sreedhar.nellaepalli@biology.ox.ac.uk.
Abstract:
The TOC translocon delivers thousands of nucleus-encoded proteins to chloroplasts and related non-photosynthetic plastids. It comprises the β-barrel channel, Toc75, and multiple isoforms of receptor GTPases, Toc33 and Toc159. However, exactly how TOC complexes are assembled in different plastid types is unknown. Here, we present detailed characterisation of two distinct TOC complexes, TOC-P and TOC-N, from photosynthetic chloroplasts and non-photosynthetic plastids, respectively. The assembled complexes are distinguished by having different sets of receptors, but both possess Toc75 which we identify as a central hub in TOC biogenesis: assembly is driven by TOC75 expression, with Toc33 and Toc159 being added sequentially thereafter. Integrative structural analysis reveals a modular architecture for TOC-P comprising a cytosolic GTPase receptor module linked flexibly to a membrane β-barrel channel module. TOC-N has a similar overall architecture, albeit with some clear differences that likely account for observed functional differences related to client specificity.
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