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Updated: May 6, 2026

Preparation of Chloroplast Sub-compartments from Arabidopsis for the Analysis of Protein Localization by Immunoblotting or Proteomics
Published on: October 19, 2018
Outer envelope membranes from chloroplasts are isolated as right-side-out vesicles
K Waegemann1, S Eichacker, J Soll
1Botanisches Institut, Universität Kiel, Olshausenstrasse 40, W-2300, Kiel, Germany.
Abstract:
Outer envelope membranes were isolated from purified chloroplasts of pea leaves. The sidedness of the vesicles was analyzed by (i) aqueous polymer-two phase partitioning, (ii) the effect of limited proteolysis on the outer-envelope proteins (OEP) 86 and OEP 7 in intact organelles and isolated membranes, (iii) fluorescence-microscopy and finally (iv) binding of precursor polypeptides to isolated outer-membrane vesicles. The results demonstrate that purified outer envelope membranes occur largely (>90%) as right-side-out vesicles.
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