Site-directed unnatural amino acid mutagenesis to investigate potassium channel pharmacology in xenopus laevis oocytes

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Abstract

Unnatural amino acid mutagenesis is a useful tool enabling detailed investigation of ion channel structure-function relationships and pharmacology. Methods have been developed to apply this technique to different heterologous systems for electrophysiological studies, with each system offering unique advantages and limitations. Synthesis of aminoacylated-tRNA followed by injection into Xenopus laevis oocytes is beneficial because it allows for the incorporation of a wide range of unnatural sidechains, including amino acids with subtle structural differences. Here, we describe a protocol for unnatural amino acid mutagenesis implemented in our lab to study the pharmacology of KCNQ voltage-gated potassium channel opener compounds. This protocol should be applicable to other ion channels and receptor types amenable for functional studies in Xenopus laevis oocytes.

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Kim, R. Y., & Kurata, H. T. (2018). Site-directed unnatural amino acid mutagenesis to investigate potassium channel pharmacology in xenopus laevis oocytes. In Methods in Molecular Biology (Vol. 1684, pp. 253–263). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7362-0_19

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