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Membrane Protein Insertion in E. coli

  • Jijun Yuan
  • Ross E. Dalbey
  • Andreas Kuhn
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 619)

Abstract

Integral membrane proteins typically span the lipid bilayer with hydrophobic α helices. These proteins can span the membrane once or multiple times with hydrophilic domains facing both sides of the membrane. In Escherichia coli, the insertion of proteins into the membrane is catalyzed by the Sec translocase and the YidC membrane insertase. YidC can function on its own to insert proteins or together with the Sec translocase to facilitate membrane protein insertion. In this chapter, we will describe the construction of a YidC depletion strain that can be used to examine whether YidC is required for membrane protein insertion. We will also present assays for determining whether a region of a membrane protein is inserted across the membrane.

Key words

YidC protease mapping AMS gel shift assay membrane protein insertion 

Notes

Acknowledgments

We thank Dr. Sandra Facey for providing Figure 4.4. This work was supported by National Institute of Health Grant (GM63862-05) to R. E. D. and the DFG grant Ku749/6-1 to A. K.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Jijun Yuan
    • 1
  • Ross E. Dalbey
    • 1
  • Andreas Kuhn
    • 2
  1. 1.Department of ChemistryThe Ohio State UniversityColumbusUSA
  2. 2.Institute of Microbiology and Molecular BiologyUniversity of HohenheimStuttgartGermany

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