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Biosynthesis of a Highly Stable Coiled-Coil Protein Containing Hexafluoroleucine in an Engineered Bacterial Host

Tang, Yi and Tirrell, David A. (2001) Biosynthesis of a Highly Stable Coiled-Coil Protein Containing Hexafluoroleucine in an Engineered Bacterial Host. Journal of the American Chemical Society, 123 (44). pp. 11089-11090. ISSN 0002-7863. https://resolver.caltech.edu/CaltechAUTHORS:20150116-154847107

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Abstract

Incorporation of nonnatural amino acid residues allows engineering of proteins with novel chemical functionality and unusual physical properties. We have shown recently that coiled-coil proteins prepared in vivo can be stabilized significantly by replacement of leucine by trifluoroleucine (1). In the same series of experiments, however, we were unsuccessful in our attempts to incorporate the more highly fluorinated analogue hexafluoroleucine (2). We report here that modification of the leucyl-tRNA synthetase (LeuRS) activity of the host allows efficient incorporation of 2 into recombinant proteins prepared in Escherichia coli. Furthermore, the coiled-coil protein used to demonstrate incorporation of 2 exhibits enhanced stability in comparison to the same protein enriched in 1, possibly due to the increased hydrophobic character of the additional trifluoromethyl group in the protein core.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/ja016652kDOIArticle
http://pubs.acs.org/doi/abs/10.1021/ja016652kPublisherArticle
http://pubs.acs.org/doi/suppl/10.1021/ja016652kPublisherSupplemental material
ORCID:
AuthorORCID
Tirrell, David A.0000-0003-3175-4596
Additional Information:Copyright © 2001 American Chemical Society. Published In Issue November 07, 2001. Publication Date (Web): October 11, 2001. Received July 18, 2001. Acknowledgment. This work was supported by the NSF MRSEC program at Caltech. Y.T. is supported by a fellowship from the Whitaker Foundation. We thank Dr. Giovanna Ghirlanda and Prof. William DeGrado for ultracentrifugation studies. We also thank Kent Kirshenbaum, Isaac Carrico, and Pin Wang for insightful discussions. Supporting Information Available: Amino acid synthesis, plasmid construction, protein expression, and protein analysis (PDF).
Funders:
Funding AgencyGrant Number
NSFUNSPECIFIED
Whitaker FoundationUNSPECIFIED
Issue or Number:44
Record Number:CaltechAUTHORS:20150116-154847107
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20150116-154847107
Official Citation:Biosynthesis of a Highly Stable Coiled-Coil Protein Containing Hexafluoroleucine in an Engineered Bacterial Host Yi Tang and and David A. Tirrell Journal of the American Chemical Society 2001 123 (44), 11089-11090
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:53848
Collection:CaltechAUTHORS
Deposited By: Katherine Johnson
Deposited On:16 Jan 2015 23:55
Last Modified:03 Oct 2019 07:53

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