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Novel Chemical Tools to Study Ion Channel Biology

Volume 869 of the series Advances in Experimental Medicine and Biology pp 119-151

Date:

Incorporation of Non-Canonical Amino Acids

  • Lilia LeisleAffiliated withDepartment of Molecular Physiology and Biophysics, University of Iowa
  • , Francis ValiyaveetilAffiliated withDepartment of Physiology and Pharmacology, Oregon Health and Sciences University
  • , Ryan A. MehlAffiliated withDepartment of Biochemistry and Biophysics, Oregon State University Corvallis
  • , Christopher A. AhernAffiliated withDepartment of Molecular Physiology and Biophysics, University of Iowa Email author 

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Abstract

In this chapter we discuss the strengths, caveats and technical considerations of three approaches for reprogramming the chemical composition of selected amino acids within a membrane protein. In vivo nonsense suppression in the Xenopus laevis oocyte, evolved orthogonal tRNA and aminoacyl-tRNA synthetase pairs and protein ligation for biochemical production of semisynthetic proteins have been used successfully for ion channel and receptor studies. The level of difficulty for the application of each approach ranges from trivial to technically demanding, yet all have untapped potential in their application to membrane proteins.

Keywords

Nonsense suppression Evolved tRNA synthetase pairs Intein ligation Protein semi-synthesis