Biosynthesis of a Highly Stable Coiled-Coil Protein Containing Hexafluoroleucine in an Engineered Bacterial Host
- Creators
- Tang, Yi
- Tirrell, David A.
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.
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).Attached Files
Supplemental Material - ja016652k_s1.pdf
Files
Name | Size | Download all |
---|---|---|
md5:bdacb2376d53840852460700d9850160
|
279.7 kB | Preview Download |
Additional details
- Eprint ID
- 53848
- Resolver ID
- CaltechAUTHORS:20150116-154847107
- NSF
- Whitaker Foundation
- Created
-
2015-01-16Created from EPrint's datestamp field
- Updated
-
2021-11-10Created from EPrint's last_modified field