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Residue-Specific Incorporation of Noncanonical Amino Acids for Protein Engineering

van Eldijk, Mark B. and van Hest, Jan C. M. (2018) Residue-Specific Incorporation of Noncanonical Amino Acids for Protein Engineering. In: Noncanonical Amino Acids. Methods in Molecular Biology. No.1728. Humana Press , New York, NY, pp. 137-145. ISBN 978-1-4939-7573-0. http://resolver.caltech.edu/CaltechAUTHORS:20180212-075402348

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Abstract

The incorporation of noncanonical amino acids has given protein chemists access to an expanded repertoire of amino acids. This methodology has significantly broadened the scope of protein engineering allowing introduction of amino acids with non-native functionalities, such as bioorthogonal reactive handles (azides and alkynes) and hydrophobic fluorinated side chains. Here, we describe the efficient residue-specific replacement of methionine by azidonorleucine in an engineered green fluorescent protein using a bacterial expression system to introduce a single reactive site for the strain-promoted azide-alkyne cycloaddition.


Item Type:Book Section
Related URLs:
URLURL TypeDescription
https://doi.org/10.1007/978-1-4939-7574-7_8DOIArticle
https://link.springer.com/protocol/10.1007%2F978-1-4939-7574-7_8PublisherArticle
Additional Information:© 2018 Springer Science+Business Media, LLC. This work was financially supported by Netherlands Organisation for Scientific Research (Rubicon grant 680-50-1407 and Gravitation program 024.001.035). The plasmids encoding the mutant MetRS and GFP were a kind gift from Prof. D. A. Tirrell (California Institute of Technology).
Funders:
Funding AgencyGrant Number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)680-50-1407
Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO)024.001.035
Subject Keywords:Noncanonical amino acids; Residue-specific incorporation; Aminoacyl-tRNA synthetase; Azidonorleucine; Medium shift; Bioconjugation; Strain-promoted azide-alkyne cycloaddition; Unnatural amino acids
Record Number:CaltechAUTHORS:20180212-075402348
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:20180212-075402348
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:84776
Collection:CaltechAUTHORS
Deposited By: Tony Diaz
Deposited On:13 Feb 2018 18:31
Last Modified:13 Feb 2018 18:31

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