Isolation of Inner and Outer Membranes of the Chloroplast Envelope from Spinach and Pea
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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.
Key words
Chloroplast Chloroplast envelope Membrane Outer envelope membrane Inner envelope membrane Differential centrifugationNotes
Acknowledgments
This work was supported by the French National Research Agency (ANR-13-ADAP-0008 Reglisse and ANR-10-LABEX-04 GRAL Labex, Grenoble Alliance for Integrated Structural Cell Biology).
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