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Heterologous Expression of Membrane Proteins for Structural Analysis

  • Isabelle Mus-VeteauEmail author
Protocol
Part of the Methods in Molecular Biology™ book series (MIMB, volume 601)

Abstract

Membrane proteins (MPs) are responsible for the interface between the exterior and the interior of the cell. These proteins are involved in numerous diseases, like cancer, cystic fibrosis, epilepsy, hyperinsulinism, heart failure, hypertension and Alzheimer disease. However, studies of these disorders are hampered by a lack of structural information about the proteins involved. Structural analysis requires large quantities of pure and active proteins. The majority of medically and pharmaceutically relevant MPs are present in tissues at low concentration, which makes heterologous expression in large-scale production-adapted cells a prerequisite for structural studies. Obtaining mammalian MP structural data depends on the development of methods that allow the production of large quantities of MPs. This review focuses on the heterologous expression systems now available to produce large amounts of MPs for structural proteomics, and describes the strategies that allowed the determination of the structure of the first heterologously expressed mammalian MPs.

Key words

Integral membrane proteins heterologous expression systems solubilization stabilization crystallization 3D structure 

Notes

Acknowledgments

I thank M. Bidet, O. Joubert and R. Nehmé for critical reading of the manuscript and the European Commission through the FP6 specific targeted research project Innovative Tools for Membrane Structural Proteomics (LSH-2003-1.1.0-1) that financially supported several authors of this volume for their research on heterologous expression systems, new surfactant molecules and MP stabilization.

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Copyright information

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

Authors and Affiliations

  1. 1.Institut of Developmental Biology and Cancer, UMR CNRS 6543, Université de Nice-Sophia Antipolis, Parc ValroseNiceFrance

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