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Mammalian Membrane Receptors Expression as Inclusion Bodies in Escherichia coli

  • Bernard Mouillac
  • Jean-Louis BanèresEmail author
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 601)

Abstract

Integral membrane proteins, in particular receptors, regulate numerous physiological functions. The primary difficulty presented by their study in vitro is to obtain them in sufficient amounts in a functional state. Escherichia coli is a host of choice for producing recombinant proteins for structural studies. However, insertion of G-protein coupled receptors into its plasma membrane usually results in bacterial death. An alternative approach consists of targeting recombinant receptors to inclusion bodies, where they accumulate without affecting bacterial growth, and then fold them in vitro . We describe here a general approach that consists of accumulating the receptor in bacterial inclusion bodies, then purifying it under denaturing conditions. A simple assay is then described to screen for refolding conditions of the protein.

Key words

Escherichia coli GPCR inclusion bodies membrane protein receptor refolding 

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Institut de Génomique FonctionnelleMontpellierFrance
  2. 2.INSERM U661MontpellierFrance
  3. 3.Université Montpellier IMontpellierFrance
  4. 4.Université Montpellier IIMontpellierFrance
  5. 5.Institut des Biomolécules Max Mousseron, Faculté de Pharmacie, Université Montpellier I, Université Montpellier IIMontpellierFrance

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