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.
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References
Heldt HW, Sauer F (1971) The inner membrane of the chloroplast envelope as the site of specific metabolite transport. Biochim Biophys Acta 234:83–91
Block MA, Dorne A-J, Joyard J, Douce R (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts I. Electrophoretic and immunochemical analyses. J Biol Chem 258:13253–13280
Cline K, Andrews J, Mersey B, Newcomb EH, Keegstra K (1981) Separation and characterization of inner and outer envelope membranes of pea chloroplasts. Proc Natl Acad Sci U S A 78:3595–3599
Arnon DI (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24:1–15
Tietje C, Heinz E (1998) Uridine-diphospho-sulfoquinovose: diacylglycerol sulfoquinovosyltransferase activity is concentrated in the inner membrane of chloroplast envelopes. Planta 206:72–78
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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Block, M.A. (2018). Isolation of Inner and Outer Membranes of the Chloroplast Envelope from Spinach and Pea. In: Maréchal, E. (eds) Plastids. Methods in Molecular Biology, vol 1829. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8654-5_9
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DOI: https://doi.org/10.1007/978-1-4939-8654-5_9
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-8654-5
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