Expression, Biochemistry, and Stabilization with Camel Antibodies of Membrane Proteins: Case Study of the Mouse 5-HT3 Receptor
There is growing interest in the use of mammalian protein expression systems, and in the use of antibody-derived 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 cryo-electron microscopy projects.
Key wordsMembrane protein Stable cell line VHH Lama antibody Cys-loop receptor
The work was supported by a grant from the European Research Council (ERC-2014-StG PentaBrain), by the Swiss National Science Foundation, by the Ecole Polytechnique Fédérale de Lausanne, and by the CEA.