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There is growing interest in the use of mammalian protein expression systems, and in the use of antibodyderived chaperones, for structural studies. Here, we describe protocols ranging from the production of recombinant membrane proteins in stable inducible cell lines to biophysical characterization of purified membrane proteins in complex with llama antibody domains. These protocols were used to solve the structure of the mouse 5-HT3 serotonin receptor but are of broad applicability for crystallization or cryoelectron microscopy projects.
Christof Holliger, Julien Maillard, Mathilde Stéphanie Willemin, Lorenzo Cimmino