Abstract
A protocol is described using lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by the substituted-cysteine accessibility method as applied to transmembrane domains (SCAM™). SCAM™ is adapted to follow changes in membrane protein topology as a function of changes in membrane lipid composition. The strategy described can be adapted to any membrane system.
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Acknowledgements
This work was supported by NIH grant GM20478 and funds from the John S. Dunn Foundation awarded to W. D.
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Bogdanov, M., Heacock, P.N., Dowhan, W. (2010). Study of Polytopic Membrane Protein Topological Organization as a Function of Membrane Lipid Composition. In: Economou, A. (eds) Protein Secretion. Methods in Molecular Biology, vol 619. Humana Press. https://doi.org/10.1007/978-1-60327-412-8_5
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DOI: https://doi.org/10.1007/978-1-60327-412-8_5
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