Abstract
Here we describe the procedures used in our laboratory for the in vitro investigation of the apparent folding kinetics as well as the folding efficiencies of outer membrane proteins (OMPs). Because microbial OMPs display a change in their gel migration upon folding, the usage of traditional gel electrophoresis is a standard method of folding analysis. Additional aspects of the method we detail herein include the preparation and storage of OMP stocks, the setup procedures for a folding reaction, and the analysis of fraction folded from scanned gel images.
The original version of this chapter was revised. The erratum to this chapter is available at: DOI 10.1007/978-1-4939-2871-2_23
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References
Burgess NK, Dao TP, Stanley AM et al (2008) Beta-barrel proteins that reside in the Escherichia coli outer membrane in vivo demonstrate varied folding behavior in vitro. J Biol Chem 283:26748–26758
Gessmann D, Chung YH, Danoff EJ et al (2014) Outer membrane β-barrel protein folding is physically controlled by periplasmic lipid head groups and BamA. Proc Natl Acad Sci U S A 111:5878–5883
Acknowledgements
This work was supported by grants from the National Science Foundation (MCB0919868), the National Institutes of Health (R01 GM079440, T32 GM008403) to KGF and NSF Fellowship DGE-1232825 to AMP.
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Plummer, A.M., Gessmann, D., Fleming, K.G. (2015). The Role of a Destabilized Membrane for OMP Insertion. In: Buchanan, S., Noinaj, N. (eds) The BAM Complex. Methods in Molecular Biology, vol 1329. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2871-2_5
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DOI: https://doi.org/10.1007/978-1-4939-2871-2_5
Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-2871-2
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