Abstract
Potassium channels conduct K+ ions selectively and at very high rates. Central to the function of K+ channels is a structural unit called the selectivity filter. In the selectivity filter, a row of four K+ binding sites are created using mainly the backbone carbonyl oxygen atoms. Due to the involvement of the protein backbone, site-directed mutagenesis is of limited utility in investigating the selectivity filter. In order to overcome this limitation, we have developed a semisynthetic approach, which permits the use of chemical synthesis to manipulate the selectivity filter. In this chapter, we describe the protocols that we have developed for the semisynthesis of the K+ channel, KcsA. We anticipate that the protocols described in this chapter will also be applicable for the semisynthesis of other integral membrane proteins of interest.
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Acknowledgements
This research was supported by grants to FIV from the NIH (GM087546), a Scientist Development Grant from the American Heart Association (0835166N) and a Pew Scholar Award.
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Komarov, A.G., Costantino, C.A., Valiyaveetil, F.I. (2013). Engineering K+ Channels Using Semisynthesis. In: Banghart, M. (eds) Chemical Neurobiology. Methods in Molecular Biology, vol 995. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-345-9_1
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DOI: https://doi.org/10.1007/978-1-62703-345-9_1
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