Membrane Protein Production in E. coli Lysates in Presence of Preassembled Nanodiscs

  • Ralf-Bernhardt Rues
  • Alexander Gräwe
  • Erik Henrich
  • Frank Bernhard
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1586)

Abstract

Cell-free expression allows to synthesize membrane proteins in completely new formats that can relatively easily be customized for particular applications. Amphiphilic superstructures such as micelles, lipomicelles, or nanodiscs can be provided as nano-devices for the solubilization of membrane proteins. Defined empty bilayers in the form of nanodiscs offer native like environments for membrane proteins, supporting functional folding, proper oligomeric assembly as well as stability. Even very difficult and detergent-sensitive membrane proteins can be addressed by the combination of nanodisc technology with efficient cell-free expression systems as the direct co-translational insertion of nascent membrane proteins into supplied preassembled nanodiscs is possible. This chapter provides updated protocols for the synthesis of membrane proteins in presence of preassembled nanodiscs suitable for emerging applications such as screening of lipid effects on membrane protein function and the modulation of oligomeric complex formation.

Key words

G-protein coupled receptors Nanodiscs Synthetic biology Membranes Oligomerization Lipid screening 

Notes

Acknowledgments

This work was funded by the Collaborative Research Center (SFB) 807 of the German Research Foundation (DFG). Support was further obtained by Instruct, part of the European Strategy Forum on Research Infrastructures (ESFRI).

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Ralf-Bernhardt Rues
    • 1
  • Alexander Gräwe
    • 1
  • Erik Henrich
    • 1
  • Frank Bernhard
    • 1
  1. 1.Centre for Biomolecular Magnetic ResonanceInstitute for Biophysical Chemistry, Goethe-University of Frankfurt/MainFrankfurt/MainGermany

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