Welcome to the new version of CaltechAUTHORS. Login is currently restricted to library staff. If you notice any issues, please email coda@library.caltech.edu
Published August 18, 2014 | public
Journal Article

In Vivo Incorporation of Non-canonical Amino Acids by Using the Chemical Aminoacylation Strategy: A Broadly Applicable Mechanistic Tool

Abstract

We describe a strategy for incorporating non-canonical amino acids site-specifically into proteins expressed in living cells, involving organic synthesis to chemically aminoacylate a suppressor tRNA, protein expression in Xenopus oocytes, and monitoring protein function, primarily by electrophysiology. With this protocol, a very wide range of non-canonical amino acids can be employed, allowing both systematic structure–function studies and the incorporation of reactive functionalities. Here, we present an overview of the methodology and examples meant to illustrate the versatility and power of the method as a tool for investigating protein structure and function.

Additional Information

© 2014 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim. Received: March 7, 2014. Article first published online: 2 July 2014. Our work on non-canonical amino acid mutagenesis has primarily been supported by the NIH (NS 34407).

Additional details

Created:
August 22, 2023
Modified:
October 26, 2023