Summary
Integral membrane proteins are a major challenge within structural genomics. These proteins are not only difficult to produce in quantities sufficient for analysis by X-ray diffraction or NMR, but also require extraction from their lipid environment, which leads to a new dimension of difficulties in purification and subsequent structural analysis. To overcome these problems requires new strategies enabling screening larger number of parameters dealing with expression and purification. For this reason, we have developed high-throughput methods for screening extracting and purifying detergents as well as other purification parameters, e.g. salt and pH. The method requires standard laboratory equipments, but can also be automated.
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Acknowledgments
The author would like to thank Professor Pär Nordlund for his advice and encouragement, Dr. Marina Ignatushchenko Sabet for her useful tips and Marie Hedrén and Victoria Lieu for technical assistance. This work was supported by the European Membrane Protein Consortium (E-MeP).
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Eshaghi, S. (2009). High-Throughput Expression and Detergent Screening of Integral Membrane Proteins. In: Doyle, S.A. (eds) High Throughput Protein Expression and Purification. Methods in Molecular Biology, vol 498. Humana Press. https://doi.org/10.1007/978-1-59745-196-3_17
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DOI: https://doi.org/10.1007/978-1-59745-196-3_17
Publisher Name: Humana Press
Print ISBN: 978-1-58829-879-9
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