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Site specific genetic incorporation of azidophenylalanine in Schizosaccharomyces pombe
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Shao, Nan, Singh, N. Sadanands, Slade, Susan E., Jones, Alexandra M. and Balasubramanian, Mohan K. (2015) Site specific genetic incorporation of azidophenylalanine in Schizosaccharomyces pombe. Scientific Reports, 5 . pp. 1-9. 17196. doi:10.1038/srep17196
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Official URL: http://dx.doi.org/10.1038/srep17196
Abstract
The diversity of protein functions is impacted in significant part by the chemical properties of the twenty amino acids, which are used as building blocks for nearly all proteins. The ability to incorporate unnatural amino acids (UAA) into proteins in a site specific manner can vastly expand the repertoire of protein functions and also allows detailed analysis of protein function. In recent years UAAs have been incorporated in a site-specific manner into proteins in a number of organisms. In nearly all cases, the amber codon is used as a sense codon, and an orthogonal tRNA/aminoacyl-tRNA synthetase (RS) pair is used to generate amber suppressing tRNAs charged with the UAA. In this work, we have developed tools to incorporate the cross-linking amino acid azido-phenylalanine (AzF) through the use of bacterial tRNATyr and a modified version of TyrRS, AzFRS, in Schizosaccharomyces pombe, which is an attractive model organism for the study of cell behavior and function. We have incorporated AzF into three different proteins. We show that the majority of AzF is modified to amino-phenyl alanine, but protein cross-linking was still observed. These studies set the stage for exploitation of this new technology for the analysis of S. pombe proteins.
| Item Type: | Journal Article | ||||||||
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| Subjects: | Q Science > QP Physiology | ||||||||
| Divisions: | Faculty of Medicine > Warwick Medical School > Biomedical Sciences > Cell & Developmental Biology Faculty of Medicine > Warwick Medical School |
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| Library of Congress Subject Headings (LCSH): | Amino acids -- Analysis | ||||||||
| Journal or Publication Title: | Scientific Reports | ||||||||
| Publisher: | Nature Publishing Group | ||||||||
| ISSN: | 2045-2322 | ||||||||
| Official Date: | 24 November 2015 | ||||||||
| Dates: |
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| Volume: | 5 | ||||||||
| Number of Pages: | 9 | ||||||||
| Page Range: | pp. 1-9 | ||||||||
| Article Number: | 17196 | ||||||||
| DOI: | 10.1038/srep17196 | ||||||||
| Status: | Peer Reviewed | ||||||||
| Publication Status: | Published | ||||||||
| Access rights to Published version: | Open Access | ||||||||
| Funder: | National University of Singapore, Wellcome Trust (London, England), Royal Society (Great Britain), University of Warwick | ||||||||
| Grant number: | WT101885MA (WT) |
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