Abstract
The study of most membrane proteins remains challenging due to their hydrophobicity and their low natural abundance in cells. Lactococcus lactis, a Gram-positive lactic bacterium, has been traditionally used in food fermentations and is nowadays widely used in biotechnology for large-scale production of heterologous proteins. This system has been successfully used for the production of prokaryotic and eukaryotic membrane proteins. The purpose of this chapter is to provide detailed protocols for (1) the expression of plant peripheral or intrinsic membrane proteins and then for (2) their solubilization, from Lactococcus membranes, for further purification steps and biochemical characterization.
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Acknowledgment
This study received financial support from the Commissariat à l’Energie Atomique et aux Energies Alternatives (CEA-PM project, postdoc fellowship to AFB), the Centre National de la Recherche Scientifique (PhD fellowship to SB), the Institut National de la Recherche Agronomique, and the University of Grenoble. ES is funded by a joint grant of the Agence Nationale de la Recherche Labex GRAL (Alliance Grenobloise pour la Biologie Structurale et Cellulaire Intégrées: ANR-10-LABEX-04) and the CEA.
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Boutigny, S. et al. (2015). Functional Expression of Plant Membrane Proteins in Lactococcus lactis . In: García-Fruitós, E. (eds) Insoluble Proteins. Methods in Molecular Biology, vol 1258. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2205-5_8
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DOI: https://doi.org/10.1007/978-1-4939-2205-5_8
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