Chapter title |
Site-Directed Unnatural Amino Acid Mutagenesis to Investigate Potassium Channel Pharmacology in Xenopus laevis Oocytes
|
---|---|
Chapter number | 19 |
Book title |
Potassium Channels
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7362-0_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7361-3, 978-1-4939-7362-0
|
Authors |
Robin Y. Kim, Harley T. Kurata |
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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