Isolation of Inner and Outer Membranes of the Chloroplast Envelope from Spinach and Pea

Abstract

The outer membrane and the inner membrane of the chloroplast envelope, also called OEM and IEM, have distinct functions connected with chloroplast biogenesis and chloroplast communication with the rest of the cell. Here we describe a method for the isolation of these membranes starting from intact chloroplast preparations, with two alternative procedures based on the starting material. One was developed from spinach leaves, the other one from pea leaves. The two procedures differ by the means that are used to isolate and rupture chloroplasts and to liberate each membrane.

Keywords: Chloroplast; Chloroplast envelope; Differential centrifugation; Inner envelope membrane; Membrane; Outer envelope membrane.