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First published online 15 January 2003
doi: 10.1242/jcs.00291

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Journal of Cell Science 116, 837-846 (2003)
doi: 10.1242/jcs.00291

Research Article

Membrane insertion of the chloroplast outer envelope protein, Toc34: constrains for insertion and topology

Soumya Qbadou1, Roselynn Tien2, Jürgen Soll1,* and Enrico Schleiff1

1 Department of Botany, Ludwig-Maximilian University Munich, 80368 Munich, Germany
2 Department of Botany, University Kiel, 24098 Kiel, Germany

* Author for correspondence (e-mail: soll{at}uni-muenchen.de)

Accepted 25 November 2002

The insertion of the outer envelope protein Toc34 from chloroplasts was studied. Toc34 was chosen as a model protein because it contains one predicted transmembrane helix at the C-terminus and a large hydrophilic N-terminal located GTPase domain, which is exposed to the cytosol. Unlike proteins located in internal chloroplast compartments, Toc34 neither contains a cleavable presequence nor uses the general import pathway. The protein can insert into the outer envelope of chloroplasts but not into the outer membrane of mitochondria. Using protein-free liposomes we showed that Toc34 is able to insert directly into the lipid bilayer. This insertion is stimulated by GTP and the presence of nonbilayer lipids, but is independent of the presence or absence of charged lipids. The topology of the protein inserted into protein-free liposomes was not exclusively directed by the positive-inside rule but by the size of the hydrophilic domain.

Key words: Direct insertion, Protein-free liposomes, Chloroplast import




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