• Open Access

Revolutionizing membrane protein overexpression in bacteria

Authors

  • Susan Schlegel,

    1. Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
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    • Contributed equally.

  • Mirjam Klepsch,

    1. Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
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    • Contributed equally.

  • Dimitra Gialama,

    1. Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
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  • David Wickström,

    1. Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
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  • Dirk Jan Slotboom,

    1. Department of Biochemistry, University of Groningen, Nyenborg 4, 9747 AG Groningen, the Netherlands.
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  • Jan-Willem De Gier

    Corresponding author
    1. Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
      E-mail: degier@dbb.su.se; Tel. (+46) 8 162420/(+46) 8 164389; Fax (+46) 8 15 3679.
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E-mail: degier@dbb.su.se; Tel. (+46) 8 162420/(+46) 8 164389; Fax (+46) 8 15 3679.

Summary

The bacterium Escherichia coli is the most widely used expression host for overexpression trials of membrane proteins. Usually, different strains, culture conditions and expression regimes are screened for to identify the optimal overexpression strategy. However, yields are often not satisfactory, especially for eukaryotic membrane proteins. This has initiated a revolution of membrane protein overexpression in bacteria. Recent studies have shown that it is feasible to (i) engineer or select for E. coli strains with strongly improved membrane protein overexpression characteristics, (ii) use bacteria other than E. coli for the expression of membrane proteins, (iii) engineer or select for membrane protein variants that retain functionality but express better than the wild-type protein, and (iv) express membrane proteins using E. coli-based cell-free systems.

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