Cell-Free Production of Membrane Proteins in Escherichia coli Lysates for Functional and Structural Studies
| Autor: | Erik Henrich, Frank Bernhard, Ralf-Bernhardt Rues, Coilin Boland, Martin Caffrey |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
030102 biochemistry & molecular biology Chemistry medicine.disease_cause Micelle Molecular biology Cell-free system 03 medical and health sciences Synthetic biology 030104 developmental biology Membrane Membrane protein medicine Biophysics Protein folding Escherichia coli G protein-coupled receptor |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493936359 |
| DOI: | 10.1007/978-1-4939-3637-3_1 |
| Popis: | The complexity of membrane protein synthesis is largely reduced in cell-free systems and it results into high success rates of target expression. Protocols for the preparation of bacterial lysates have been optimized in order to ensure reliable efficiencies in membrane protein production that are even sufficient for structural applications. The open accessibility of the semisynthetic cell-free expression reactions allows to adjust membrane protein solubilization conditions according to the optimal folding requirements of individual targets. Two basic strategies will be exemplified. The post-translational solubilization of membrane proteins in detergent micelles is most straightforward for crystallization approaches. The co-translational integration of membrane proteins into preformed nanodiscs will enable their functional characterization in a variety of natural lipid environments. |
| Databáze: | OpenAIRE |
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