© Springer Science+Business Media New York 2015 119
C. Ahern, S. Pless (eds.), Novel Chemical Tools to Study Ion Channel Biology,
Advances in Experimental Medicine and Biology 869,
DOI 10.1007/978-1-4939-2845-3_7
C. A. Ahern () · L. Leisle
Department of Molecular Physiology and Biophysics, University of Iowa, 51 Newton Road,
Iowa City, IA 52246, USA
e-mail: [email protected]
F. Valiyaveetil
Department of Physiology and Pharmacology, Oregon Health and Sciences University, Portland,
OR 97239, USA
R. A. Mehl
Department of Biochemistry and Biophysics, Oregon State University Corvallis, Corvallis, OR
97331, USA
Incorporation of Non-Canonical Amino Acids
Lilia Leisle, Francis Valiyaveetil, Ryan A. Mehl and Christopher A. Ahern
Abstract In this chapter we discuss the strengths, caveats and technical consider-
ations of three approaches for reprogramming the chemical composition of selected
amino acids within a membrane protein. In vivo nonsense suppression in the Xeno-
pus 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
1 Introduction
The ability to genetically alter the chemo-physical properties of an amino acid per-
mits the detailed examination of the functional contributions of atomic character-
istics of side- and main-chain chemistries. Further, emerging techniques are avail-
able to genetically encode non-canonical amino acids (ncAA) with fluorescent/