Waegemann, Karin; Eichacker, Susanne; Soll, Jürgen
Outer envelope membranes from chloroplasts are isolated as right-side-out vesicles.
In: Planta, Vol. 187, No. 1: pp. 89-94
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.